Human Parvovirus: Bocavirus

Abstract

Provided herein are sequences of the genomes and encoded proteins of a new human parvovirus, Bocavirus-2, and variants thereof. Also provided are methods of detecting the Bocavirus-2 and diagnosing Bocavirus-2 infection, methods of treating or preventing Bocavirus-2 infection, and methods for identifying anti-Bocavirus-2 compounds.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation application of U.S. application Ser. No. 12/426,173 filed Apr. 17, 2009, now issued as U.S. Pat. No. 8,071,751; which claims the benefit under 35 USC §119(e) to U.S. Application Ser. No. 60/225,264 filed Aug. 15, 2000, now expired. The disclosure of each of the prior applications is considered part of and is incorporated by reference in the disclosure of this application.

BACKGROUND OF THE INVENTION

[0003] 1. Field of the Invention

[0004] The present invention relates generally to the discovery of a new human parvovirus and more specifically, to methods of using the virus including methods of detecting the virus and diagnosing viral infection, methods of treating or preventing virus infection, and methods for identifying anti-viral compounds.

[0005] 2. Background of the Invention

[0006] Parvoviruses are among the smallest DNA-containing viruses that infect animals and man. Parvoviruses range in size from 15 to 28 nm in diameter, lack a lipid membrane (non-enveloped), and contain a single strand of DNA. Parvoviruses are heat stable and generally resistant to chemical deactivating, agents, which may account for their prevalence and persistence in the environment. In animals, many diseases such as canine parvovirus and feline panleukopenia exhibit high morbidity and high mortality in affected animal populations and the infections can persist endemically.

[0007] The Parvoviridae family is divided into five genera: Parvovirus, Dependovirus, Erythrovirus, Amdovirus and Bocavirus. Animal parvoviruses such as canine parvovirus, feline parvovirus, mink enteritis virus, and porcine parvovirus are responsible for many serious diseases in animals. In humans, the first identified pathogenic member of this family was parvovirus B19, which is a member of genus Erythrovirus. Other B19-related human parvoviruses include A6 and V9. The genomes of A6 and V9 are highly related to that of B19. As with other parvoviruses, B19 is highly contagious and exhibits high morbidity in affected populations. B19 causes fifth disease in normal individuals, transient aplastic crisis in patients with underlying hemolysis, and chronic anemia due to persistent infection in immunocompromised patients. B19 infection in pregnancy can lead to hydrops fetalis and fetal loss. B19 has also been implicated as the cause of chronic arthritis in adults where there is evidence of recent B19 infection, e.g., rheumatoid and inflammatory arthritis.

[0008] Parvoviruses are also associated with respiratory tract infections. Lower respiratory tract infections (LRTI) are a leading cause of hospitalization of infants and young children. Animal bocaviruses BPV (bovine parvovirus) and MVC (canine minute virus, or minute virus of canines) are associated with respiratory symptoms and enteritis of young animals. Systemic infection by BPV and MVC appears likely, and there are indications that fetal infection leading to fetal death may occur.

[0009] The discovery of a human Bocavirus (HBoV) has been recently reported. (WO2007/057062) HBoV is pathogenic to humans, and is associatied with respiratory tract infections in children.

[0010] Despite the known pathogenicity of parvoviruses and the urgent need for methods to prevent, diagnose and treat parvovirus infections, other divergent human parvoviruses have not yet been identified. Therefore, a need exists to detect divergent human parvoviruses and to provide a method to diagnose, prevent and treat parvoviruses infection. Moreover, there exists a need to provide methods to identify parvoviruses antiviral compounds.

SUMMARY OF THE INVENTION

[0011] The present invention relates to a new human Parvovirus, Bocavirus-2 (HBoV2). Accordingly, the present invention provides the genomic sequences of 9 variants of Bocavirus-2, and the sequences of the viral proteins encoded thereby. The variants have been termed HBoV2A-PK5510, HBoV2A-PK2255, HBoV2A-UK648, HBoV2A-TU114-06, HBoV2B-NI327, HBoV2B-NI213, HBoV3-NI385, HBoV4-TU210-07, and HBoV4-NI374. Also provided are methods of detecting the Bocavirus-2 variants and diagnosing Bocavirus-2 infection in biological samples, methods of treating or preventing Bocavirus-2 infection, and methods for identifying antiviral compounds.

[0012] Accordingly, in one embodiment of the present invention there are provided isolated nucleic acid molecules obtained from Bocavirus-2 variants. In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence having at least 80% identity to SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In some embodiments, the nucleic acid molecule comprises a nucleotide sequence having at least 85% identity to SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In other embodiments, the nucleic acid molecule comprises a nucleotide sequence having at least 90% identity to SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In still other embodiments, the nucleic acid molecule comprises a nucleotide sequence having at least 95% identity to SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In one aspect, the nucleic acid molecule comprises a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, and a complement thereof. In still other embodiments, the nucleic acid molecule is a fragment of at least 12 nucleotides in length of any of the above nucleic acids. In some embodiments, the fragment may be at least 20, 25, 30, 40, 50, 75, 100, or 200 nucleotides in length.

[0013] In certain embodiments, the nucleic acid molecule comprises a nucleotide sequence that hybridizes under highly stringent conditions to at least 12 contiguous nucleotides of the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In particular embodiments, the nucleotide sequence hybridizes under stringent conditions to at least 25, or at least 50, or at least 100, or at least 150 contiguous nucleotides of the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In one aspect, the nucleotide sequence hybridizes under highly stringent conditions over the full length of the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In one aspect, the nucleic acid molecule is at least 12 nucleotides in length. In another aspect, the nucleotide sequence comprises at least 80% identity, at least 90% identity, or at least 95% identity to SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof.

[0014] In another embodiment, the nucleic acid molecule hybridizes under highly stringent conditions to at least 12 contiguous nucleotides of an open reading frame of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In one aspect, the nucleotide sequence comprises an open reading frame encoding a protein selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, and conservative variants thereof.

[0015] In another embodiment of the invention, there are provided substantially purified proteins encoded by Bocavirus-2 nucleic acid molecules of the invention. In some embodiments, the protein is encoded by a nucleic acid sequence that hybridizes under stringent conditions to at least 12 contiguous nucleotides of the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In certain embodiments, the protein is encoded by a nucleic acid sequence that hybridizes under stringent conditions to at least 25, or at least 50, or at least 100, or at least 150 contiguous nucleotides of the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof. In particular embodiments, the invention includes a protein encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, and SEQ ID NO:49. In other embodiments, the protein comprises a sequence having about 80%, or 90%, or 95% identity to a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, and fragments thereof. In some embodiments, the protein comprises a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, and fragments thereof. In certain embodiments the fragment is an antigen or immunogenic fragment.

[0016] In one embodiment, the invention includes a composition containing a protein of the invention.

[0017] The invention also provides an isolated antibody that specifically binds to a protein of the invention. In one aspect, the antibody is a polyclonal antibody. In another aspect, the antibody is a monoclonal antibody.

[0018] In another embodiment, the invention includes purified serum containing polyclonal antibodies that specifically bind to a protein of the invention.

[0019] The invention also provides an isolated Bocavirus-2 comprising a nucleic acid molecule of the invention.

[0020] In one embodiment, there is provided an expression vector comprising a nucleic acid molecule of the invention. In one aspect, a host cell comprising the expression vector is provided.

[0021] In another embodiment, the invention includes a substantially pure preparation of virus which induces respiratory tract infection.

[0022] In still another embodiment of the invention, there is provided a method of detecting a Bocavirus-2 nucleic acid molecule by hybridization to a probe. In some embodiments, the method includes contacting, under highly stringent hybridization conditions, a sample suspected of containing a Bocavirus-2 nucleic acid with a nucleotide sequence that hybridizes under highly stringent conditions to a nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof; and detecting the presence or absence of hybridization. In one aspect, the hybridization conditions include hybridizing at 42° C. in a solution comprising 50% formamide, 5×SSC, and 1% SDS and washing at 65° C. in a solution comprising 0.2×SSC and 0.1% SDS.

[0023] In yet another embodiment of the invention, there is provided a method of detecting a Bocavirus-2 nucleic acid molecule by detection of a nucleic acid amplification product. In some embodiments the method includes amplifying the nucleic acid of a sample suspected of containing Bocavirus-2 nucleic acid with at least one primer that hybridizes to a nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof to produce an amplification product; and detecting the presence of an amplification product, thereby detecting the presence of the Bocavirus-2 nucleic acid. In one aspect, the amplifying includes a denaturation phase of 90° C.-95° C. for 30 sec-2 min., an annealing phase of 50° C. to about 65° C. lasting 30 sec.-2 min., and an extension phase of about 72° C. for 1-2 min., and an extension phase of about 72° C. for 1-2 min for 20-40 cycles.

[0024] In another embodiment, there is provided a method of detecting a Bocavirus-2 infection in a subject by detecting a protein of the invention in a sample from the subject. In one aspect the method includes contacting a sample suspected of comprising a Bocavirus-2 protein with an antibody that specifically binds a polypeptide encoded by SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, or SEQ ID NO:49 to form a protein/antibody complex; and detecting the presence of the protein/antibody complex, thereby detecting the presence of the Bocavirus protein.

[0025] The invention also contemplates a kit for detecting a Bocavirus-2 nucleic acid, the kit containing at least one polynucleotide having a nucleotide sequence that hybridizes under highly stringent conditions to a nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof.

[0026] The invention also contemplates a kit for detecting a Bocavirus-2 nucleic acid, the kit containing at least one oligonucleotide primer that hybridizes to a nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof under highly stringent PCR conditions. In one aspect the PCR conditions comprise a denaturation phase of 90° C.-95° C. for 30 sec-2 min., an annealing phase of 50° C. to about 65° C. lasting 30 sec.-2 min., and an extension phase of about 72° C. for 1-2 min., and an extension phase of about 72° C. for 1-2 min for 20-40 cycles.

[0027] In another embodiment, the invention describes a kit for detecting a Bocavirus-2 in a sample, where the kit contains an antibody that detects a polypeptide encoded by SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, or SEQ ID NO:49. In one aspect, the kit contains a monoclonal antibody. In another aspect, the kit contains a polyclonal antibody.

[0028] The invention contemplates a method of assaying for an anti-Bocavirus-2 compound by 1) contacting a sample suspected of containing a Bocavirus-2 with a test compound, where the Bocavirus encodes a genome that hybridizes under highly stringent conditions to a nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof, wherein the hybridization reaction is incubated at 42° C. in a solution comprising 50% formamide, 5×SSC, and 1% SDS and washed at 65° C. in a solution comprising 0.2×SSC and 0.1% SDS; and 2) determining whether the test compound inhibits Bocavirus replication, wherein inhibition of Bocavirus replication indicates that the test compound is the anti-Bocavirus compound.

[0029] In still other embodiments, there is provided a method of treating or preventing a Bocavirus-2 infection in a subject by administering to the subject an antigen encoded by a Bocavirus, the Bocavirus containing a genome that hybridizes under highly stringent conditions to a nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof; thereby treating or prevention infection in the subject.

[0030] Another embodiment of the invention provides a vaccine for the prevention of respiratory tract infections in a subject, including a Bocavirus-2 or viral antigen from a Bocavirus-2, which induces respiratory tract infections in a subject and a pharmacologically acceptable carrier. In one aspect, the virus of the vaccine is in a killed form. In another aspect, the virus of the vaccine is in a live but attenuated form.

[0031] In one embodiment of the invention, there is provided a method for detecting and serotyping Bocavirus in a sample by 1) contacting a first portion of the sample with a first pair of primers in a first amplification protocol, wherein the first pair of primers have an associated first characteristic amplification product if a Bocavirus is present in the sample; 2) determining whether or not the first characteristic amplification product is present; 3) contacting a second portion of the sample with a second pair of primers in a second amplification protocol, wherein the second pair of primers have an associated second characteristic amplification product if a Bocavirus is present in the sample and wherein the second pair of primers are different from the first pair of primers; 4) determining whether or not the second characteristic amplification product is present; 5) based on whether or not the first and second characteristic amplification product are present, selecting one or more subsequent pair of primers and contacting the one or more subsequent pair of primers with additional portions of the sample in subsequent amplification protocols, wherein each subsequent pair of primers is different from each pair of primers already used and wherein each subsequent pair of primers has an associated subsequent characteristic amplification product if a Bocavirus is present in the sample; 6) determining whether or not the associated characteristic amplification product for each subsequent pair of primers used is present; 7) repeating steps 5 and 6 for one or more subsequent pairs of primers if the Bocavirus cannot be serotyped based on the determinations of steps 2, 4, and 6 until the Bocavirus can be serotyped, wherein the one or more subsequent pairs of primers are different from all pairs of primers used in earlier amplification protocols; and determining the serotype or groups of serotypes of the Bocavirus that may be present in the sample. In one aspect, the sample is a biological sample. In another aspect, the sample is whole blood or a fraction thereof, a bronchial wash, cerebrospinal fluid, an eye swab, a conjunctival swab, a swab or scraping from a lesion, a nasopharyngeal swab, an oral or buccal swab, pericardial fluid, a rectal swab, serum, semen, cerebrospinal fluid, sputum, saliva, stool, a stool extract, a throat swab, urine, brain tissue, heart tissue, intestinal tissue, kidney tissue, liver tissue, lung tissue, pancreas tissue, spinal cord tissue, skin tissue, spleen tissue, thymus tissue, cells from a tissue culture, a supernatant from a tissue culture, and tissue from an experimentally infected animal. In one aspect, the first, second, and any subsequent amplification protocols are polymerase chain reactions or reverse-transcription polymerase chain reactions. In another aspect, detecting and serotyping of the Bocavirus in the sample is used to diagnose a viral disease or medical condition. In yet another aspect, the viral disease or medical condition is a respiratory tract infection.

[0032] In still another embodiment of the invention, there is provided a method for detecting the presence of a Bocavirus in a sample by 1) purifying RNA contained in the sample; 2) reverse transcribing the RNA with primers effective to reverse transcribe Bocavirus RNA to provide a cDNA; 3) contacting at least a portion of the cDNA with (i) a composition that promotes amplification of a nucleic acid and (ii) an oligonucleotide mixture wherein the mixture comprises at least one oligonucleotide that hybridizes to a highly conserved sequence of the sense strand of a Bocavirus nucleic acid and at least one oligonucleotide that hybridizes to a highly conserved sequence of the antisense strand of a Bocavirus nucleic acid; 4) carrying out an amplification procedure on the amplification mixture such that, if a Bocavirus is present in the sample, a Bocavirus amplicon is produced whose sequence comprises a nucleotide sequence of at least a portion of the Bocavirus genome; and 5) detecting whether an amplicon is present; wherein the presence of the amplicon indicates that a Bocavirus is present in the sample. In one aspect, the amplification procedure comprises a polymerase chain reaction. In another aspect, the sample is chosen from the group consisting of whole blood or a fraction thereof, a bronchial wash, cerebrospinal fluid, an eye swab, a conjunctival swab, a swab or scraping from a lesion, a nasopharyngeal swab, an oral or buccal swab, pericardial fluid, a rectal swab, serum, semen, cerebrospinal fluid, sputum, saliva, stool, a stool extract, a throat swab, urine, brain tissue, heart tissue, intestinal tissue, kidney tissue, liver tissue, lung tissue, pancreas tissue, spinal cord tissue, skin tissue, spleen tissue, thymus tissue, cells from a tissue culture, a supernatant from a tissue culture, and tissue from an experimentally infected animal. In another aspect, the detection is carried out by a procedure chosen from the group consisting of gel electrophoresis and visualization of amplicons contained in a resulting gel, size separation matrix, capillary electrophoresis and detection of the emerging amplicon, probing for the presence of the amplicon using a labeled probe, sequencing the amplicon, and labeling a PCR primer employed in the method and detecting the label.

BRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIG. 1 shows the nucleic acid sequence of the human Bocavirus-type-2 (HBoV2A-PK5510) genome (SEQ ID NO:1).

[0034] FIG. 2 shows the amino acid sequences of the HBoV2A-PK5510 nonstructural proteins NS1 (SEQ ID NO:2) and NS2 (SEQ ID NO:29).

[0035] FIG. 3 shows the amino acid sequence of the HBoV2A-PK5510 nucleoprotein protein (NP1) (SEQ ID NO:3).

[0036] FIG. 4 shows the amino acid sequence of the HBoV2A-PK5510 proteins VP1 (SEQ ID NO:4) and VP2 (SEQ ID NO:30).

[0037] FIG. 5 shows the nucleotide alignment between the complete genome of HBoV1 (SEQ ID NO:55) and HBoV2 (SEQ ID NO:1).

[0038] FIG. 6 shows the amino acid alignments between HBoV2 and HBoV1 for the a) NS1 protein (SEQ ID NO'S 2 & 56, respectively), b) NP1 protein (SEQ ID NO'S 3 & 57, respectively, and c) VP1 protein (SEQ ID NO'S 58 & 59, respectively).

[0039] FIGS. 7A-D show the genomic sequence of HBoV4-TU210-07 (SEQ ID NO:5; FIG. 7A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:6 and SEQ ID NO:31, respectively; FIG. 7B); NP1 (SEQ ID NO:7; FIG. 7C); and VP1 and VP2 (SEQ ID NO:8 and SEQ ID NO:32, respectively; FIG. 7D).

[0040] FIGS. 8A-D show the genomic sequence of HBoV2A-TU114-06 (SEQ ID NO:9; FIG. 8A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:10 and SEQ ID NO:33, respectively; FIG. 8B); NP1 (SEQ ID NO:11; FIG. 8C); and VP1 and VP2 (SEQ ID NO:12 and SEQ ID NO:34, respectively; FIG. 8D).

[0041] FIGS. 9A-D show the genomic sequence of HBoV2B-NI213 (SEQ ID NO:13; FIG. 9A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:14 and SEQ ID NO:35, respectively; FIG. 9B); NP1 (SEQ ID NO:15; FIG. 9C); and VP1 and VP2 (SEQ ID NO:16 and SEQ ID NO:36, respectively; FIG. 9D).

[0042] FIGS. 10A-D show the genomic sequence of HBoV2B-NI327 (SEQ ID NO:17; FIG. 10A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:18 and SEQ ID NO:37, respectively; FIG. 10B); NP1 (SEQ ID NO:19; FIG. 10C); and VP1 and VP2 (SEQ ID NO:20 and SEQ ID NO:38, respectively; FIG. 10D).

[0043] FIGS. 11A-D show the genomic sequence of HBoV4-NI374 (SEQ ID NO:21; FIG. 11A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:22 and SEQ ID NO:39, resepectively; FIG. 11B); NP1 (SEQ ID NO:23; FIG. 11C); and VP1 and VP2 (SEQ ID NO:24 and SEQ ID NO:40, respectively; FIG. 11D).

[0044] FIGS. 12A-D show the genomic sequence of HBoV3-NI3853 (SEQ ID NO:25; FIG. 12A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:26 and SEQ ID NO:41, respectively; FIG. 12B); NP1 (SEQ ID NO:27; FIG. 12C); and VP1 and VP2 (SEQ ID NO:28 and SEQ ID NO:42, respectively; FIG. 12D).

[0045] FIG. 13 shows a schematic representation of the genomic organization of HBoV2 and other species of Bocaviruses (BPV, CnMV, and HBoV).

[0046] FIG. 14 shows a phylogenetic analysis of Bocaviruses. The 3 major open reading frames were analyzed using both nucleotide and protein sequences of representative variants of HBoV, HParV5, CnMV (Canine minute virus) and BPV-1 (Bovine parvovirus-1). Analysis of the partial sequence of NS1 was used to show phylogenetic relationships of between a larger number of samples amplified by PCR; it also corresponded to the region of the partially sequenced Brazilian HBoV variant (EF560212).

[0047] FIG. 15 shows a sequence divergence scan of the complete genome sequences of Bocavirus-2 variants (gt1: HBoV2A-PK5510; gt2: HBoV2A-PK2255; gt3: HBoV2A-UK648).

[0048] FIGS. 16A-D show the genomic sequence of HBoV2A-PK2255 (SEQ ID NO:43; FIG. 16A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:44 and SEQ ID NO:45, respectively; FIG. 16B); NP1 (SEQ ID NO:46; FIG. 16C); and VP1 and VP2 (SEQ ID NO:47 and SEQ ID NO:48, respectively; FIG. 16D).

[0049] FIGS. 17A-D show the genomic sequence of HBoV2A-UK648 (SEQ ID NO:49; FIG. 17A) and the encoded amino acid sequences for NS1 and NS2 (SEQ ID NO:50 and SEQ ID NO:51, respectively; FIG. 17B); NP1 (SEQ ID NO:52; FIG. 17C); and VP1 and VP2 (SEQ ID NO:53 and SEQ ID NO:54, respectively; FIG. 17D).

DETAILED DESCRIPTION OF THE INVENTION

[0050] The present invention is based on the discovery of a novel virus, Bocavirus-2, which is associated with respiratory tract infections. There are various molecular methods for discovery of novel human viruses including immunoreactive cDNA expression library screening, representational difference analysis (RDA), DNA microarrays and use of degenerate PCR primers. Other methods include sequence independent single primer amplification of nucleic acids in serum (DNase-SISPA), or "metagenomics shotgun sequencing." For these approaches, DNA can be isolated directly from environmental samples and sequenced, without attempting to culture the organisms from which it comes. The DNase-SISPA method first removes contaminating human DNA in plasma or serum by DNase digestion. Viral nucleic acids protected from DNase digestion by their viral coats are then converted into double stranded DNA (dsDNA) using random primers. The dsDNA is then digested by a four base pair specific restriction endonuclease resulting in two overhanging bases to which are ligated a complementary oligonucleotide linker. A PCR primer complementary to the ligated linker is then used to PCR amplify the sequences between the restriction sites. The PCR products are analyzed by PAGE and distinct DNA bands are extracted, subcloned and sequenced. Similarity to known viruses is then tested using BLASTn (for nucleic acid similarity) and tBLASTx (for protein similarity). The DNase-SISPA method does not require foreknowledge of the viral sequences being amplified and can therefore theoretically amplify more divergent members of known viral families than nucleic acid sequence similarity-dependent approaches using degenerate primers or microarrays.

[0051] Accordingly, the present invention provides a new virus, human Bocavirus-2 (HBoV2) and variants thereof, as well as their genomic sequences and the viral proteins encoded thereby. The genome encodes the structural proteins of the virus and non-structural proteins involved in viral replication. The genomic sequence of an exemplary Bocavirus-2 (termed HBoV2A-PK5510) is provided herein. In addition, 8 variant Bocavirus-2 genomes are provided herein, termed HBoV2A-PK2255, HBoV2A-UK648, HBoV2A-TU114-06, HBoV2B-NI327, HBoV2B-NI213, HBoV3-NI385, HBoV4-TU210-07, and HBoV4-NI374. The genomic sequences of these bocavirus-2 variants are provided in FIG. 1 (HBoV2A-PK5510; SEQ ID NO:1), FIG. 7A (HBoV4-TU210-07; SEQ ID NO:5), FIG. 8A (HBoV2A-TU114-06; SEQ ID NO:9), FIG. 9A (HBoV2B-NI213; SEQ ID NO:13), FIG. 10A (HBoV2B-NI327; SEQ ID NO:17), FIG. 11A (HBoV4-NI374; SEQ ID NO:21), FIG. 12A (HBoV3-NI3853; SEQ ID NO:25), FIG. 16A (HBoV2A-PK2255; SEQ ID NO:43), and FIG. 17A (HBoV2A-UK648; SEQ ID NO:49).

[0052] The amino acid sequences of exemplary HBoV2 (HBoV2A-PK5510) nonstructural proteins NS1 and NS2 are shown in FIG. 2 (SEQ ID NO:2 and SEQ ID NO:29, respectively). Additional exemplary amino acid sequences of NS1 proteins are set forth in SEQ ID NOs: 6, 10, 14, 18, 22, 26, 44, and 50. Additional exemplary amino acid sequences of NS2 proteins are set forth in SEQ ID NOs: 31, 33, 35, 37, 39, 41, 45, and 51. FIG. 3 shows the amino acid sequence of an exemplary HBoV2 nucleoprotein protein (NP) (SEQ ID NO:3). Additional exemplary amino acid sequences of NP proteins are set forth in SEQ ID NOs: 7, 11, 15, 19, 23, 27, 46, and 52. Additionally, FIG. 4 shows the amino acid sequences of exemplary HBoV2 VP1 and VP2 proteins (SEQ ID NO:4 and SEQ ID NO:30, respectively). Further exemplary amino acid sequences of VP1 proteins are set forth in SEQ ID NOs: 8, 12, 16, 20, 24, 28, 47, and 53. Further exemplary amino acid sequences of VP2 proteins are set forth in SEQ ID NOs: 30, 32, 34, 36, 38, 40, 42, 48, and 54.

[0053] Phylogenetic analysis of HBoV2 establishes it as diverse from HBoV1 (also termed HBoV), containing a 78% amino acid identity to HBoV1 in the non-structural (NS) region. (FIG. 6a) The nucleocapsid protein (NP) of HBoV1 has only a 66% amino acid identity with the NP protein of HBoV-2. (FIG. 6b) Additionally, the capsid protein (CP) of HBoV has only a 66% amino acid identity to the NP protein of HBoV2. (FIG. 6c)

[0054] The identification of Bocavirus-2 provides methods of detecting the virus, its genome, transcripts, and proteins including structural and non-structural proteins. Antibodies (polyclonal and monoclonal) made to antigens from any of these viral proteins can be used to detect the antigen or protein as well as to isolate the antigens and to remove virus, proteins, or nucleic acids from a sample, e.g., a blood sample. Antibodies to Bocavirus-2 antigens can be used in diagnostic assays to detect viral infection. Any suitable sample, including blood, saliva, sputum, etc., can be used in a diagnostic assay of the invention. Such antibodies can also be used in therapeutic applications to inhibit or prevent viral infection.

[0055] The Bocavirus-2 antigens of the invention can also be used in diagnostic application to detect anti-Bocavirus-2 antigen antibodies in infected or exposed subjects. Bocavirus-2 antigens of the invention can also be used therapeutically, as prophylactic vaccines or vaccines for acute or latent infections, e.g., whole virus vaccines, protein or subunit vaccines, and nucleic acid vaccines encoding viral proteins, ORFs or genomes for intracellular expression and secretion or cell surface display, or can be targeted to specific cell types using promoters and vectors.

[0056] The Bocavirus virus, nucleic acids and proteins of the invention can be used to assay for antiviral compounds, including compounds that inhibit (1) viral interactions at the cell surface, e.g., viral transduction (e.g., block viral cell receptor binding or internalization); (2) viral replication and gene expression, e.g., viral replication (e.g., by inhibiting non-structural protein activity, origin activity, or primer binding), viral transcription (promoter or splicing inhibition, nonstructural protein inhibition), viral protein translation, protein processing (e.g., cleavage or phosphorylation); and (3) viral assembly and egress, e.g., viral packaging, and virus release.

[0057] "Bocavirus" refers to both the genetic components of the virus, e.g., the genome (positive or negative) and RNA transcripts thereof (either sense or antisense), proteins encoded by the genome (including structural and nonstructural proteins), and viral particles. This description includes Bocavirus nucleic acids, alleles, mutants, and interspecies homologs that: (1) have a nucleotide sequence that has greater than about 60% nucleotide sequence identity, 65%, 70%, 75%, 80%, 85%, 90%, preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or greater nucleotide sequence identity, preferably over a region of at least about 25, 50, 100, 200, 500, 1000, or more nucleotides, up to the full length sequence, to the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof; (2) encode a protein that binds to antibodies, e.g., polyclonal or monoclonal antibodies, raised against an antigen or an immunogen from an amino acid sequence of a protein encoded by an open reading frame of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, or SEQ ID NO:49; (3) specifically hybridize under stringent hybridization conditions to a nucleic acid sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof; (4) encodes a protein having an amino acid sequence that has greater than about 60% identity, 65%, 70%, 75%, 80%, 85%, 90%, preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or greater sequence identity, preferably over a region of over a region of at least about 25, 50, 100, 200, 500, 1000, or more amino acids, to a protein as set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, or SEQ ID NO:54. Bocavirus-2 nucleic acids may be isolated from an animal host including, but not limited to, primate, e.g., human; rodent, e.g., rat, mouse, hamster; cow, pig, horse, sheep, or any mammal. The nucleic acids and proteins of the invention include both naturally occurring and recombinant molecules.

[0058] Bocavirus-2 nucleic acids can be used to produce infectious clones, e.g., for production of recombinant viral particles, including empty capsids or capsids containing a recombinant (e.g., wild type or further comprising a heterologous nucleic acid) or modified (e.g., mutated) Bocavirus genome, which may be replication competent or incompetent, using the methods disclosed in U.S. Pat. Nos. 6,558,676; 6,132,732; 6,001,371; 5,916,563; 5,827,647; 5,508,186; 6,379,885; 6,287,815; 6,204,044; and 5,449,608. Such particles are useful as gene transfer vehicles, and as vaccines, and for use in diagnostic applications and for drug discovery assays for antiviral compounds, as discussed below.

[0059] "Protein encoded by Bocavirus" or "protein encoded by Bocavirus open reading frame (ORF)" refers to structural and non-structural Bocavirus proteins that: (1) are encoded by a nucleic acid molecule of the invention such as a nucleic acid molecule that has greater than about 60% nucleotide sequence identity, 65%, 70%, 75%, 80%, 85%, 90%, preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or greater nucleotide sequence identity, preferably over a region of over a region of at least about 25, 50, 100, 200, 500, 1000, or more nucleotides, up to the full length sequence, to the nucleotide sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof; (2) bind to antibodies, e.g., polyclonal or monoclonal antibodies, raised against an immunogen comprising an amino acid sequence of a protein as set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or conservatively modified variants thereof; (3) is encoded by a nucleic acid molecule that specifically hybridizes under stringent hybridization conditions to a nucleic acid sequence as set forth in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof, starting with an ATG and ending with a TAA, e.g., nucleotides 2378-5122); (4) have an amino acid sequence that has greater than about 60% identity, 65%, 70%, 75%, 80%, 85%, 90%, preferably 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% or greater identity, preferably over a region of at least about 25, 50, 100, 200, 500, 1000, or more amino acids, to a protein as set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, or conservatively modified variants thereof.

[0060] "Biological sample" includes sections of tissues such as biopsy and autopsy samples, and frozen sections taken for histologic purposes. Such samples include blood and blood fractions or products (e.g., serum, plasma, platelets, red blood cells, and the like), sputum, tissue, cultured cells, e.g., primary cultures, explants, and transformed cells, stool, urine, etc. A biological sample is typically obtained from a eukaryotic organism, most preferably a mammal such as a primate e.g., chimpanzee or human; cow; dog; cat; a rodent, e.g., guinea pig, rat, mouse; rabbit; or a bird; reptile; or fish.

[0061] The terms "identical" or percent "identity," in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region (e.g., nucleotide sequence or amino acid sequence of FIGS. 1 and 2, respectively, corresponding to SEQ ID NO:1 and SEQ ID NO:2, respectively), when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection. Such sequences are then said to be "substantially identical." This definition also refers to, or may be applied to, the compliment of a test sequence. The definition also includes sequences that have deletions and/or additions, as well as those that have substitutions. As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 amino acids or nucleotides in length.

[0062] For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Preferably, default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.

[0063] There are several methods available and well known to those skilled in the art to obtain full-length DNAs, or extend short DNAs, for example those based on the method of Rapid Amplification of cDNA Ends (RACE). Another sequencing method is based on detecting the activity of DNA polymerase with a chemiluminescent enzyme. Essentially, the method allows sequencing of a single strand of DNA by synthesizing the complementary strand along it, one base pair at a time, and detecting which base was actually added at each step. The template DNA is immobilized, and solutions of A, C, G, and T nucleotides are added sequentially. Light is produced only when the nucleotide solution compliments the first unpaired base of the template. The sequence of solutions which produce chemiluminescent signals allows the determination of the sequence of the template.

[0064] A "comparison window," as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 supplement)).

[0065] A preferred example of algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990), respectively. BLAST and BLAST 2.0 are used, with the parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands.

[0066] "Nucleic acid" refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, and complements thereof. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).

[0067] Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues. The term nucleic acid is used interchangeably with gene, cDNA, mRNA, oligonucleotide, and polynucleotide.

[0068] A particular nucleic acid sequence also implicitly encompasses "splice variants." Similarly, a particular protein encoded by a nucleic acid implicitly encompasses any protein encoded by a splice variant of that nucleic acid. "Splice variants," as the name suggests, are products of alternative splicing of a gene. After transcription, an initial nucleic acid transcript may be spliced such that different (alternate) nucleic acid splice products encode different polypeptides. Mechanisms for the production of splice variants vary, but include alternate splicing of exons. Alternate polypeptides derived from the same nucleic acid by read-through transcription are also encompassed by this definition. Any products of a splicing reaction, including recombinant forms of the splice products, are included in this definition.

[0069] The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymer.

[0070] The term "amino acid" refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.

[0071] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.

[0072] "Conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, conservatively modified variants refers to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of skill will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence with respect to the expression product, but not with respect to actual probe sequences.

[0073] As to amino acid sequences, one of skill will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively modified variant" where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles of the invention.

[0074] The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and 8) Cysteine (C), Methionine (M).

[0075] Macromolecular structures such as polypeptide structures can be described in terms of various levels of organization. For a general discussion of this organization, see, e.g., Alberts et al., Molecular Biology of the Cell (3^rd ed., 1994) and Cantor and Schimmel, Biophysical Chemistry Part I: The Conformation of Biological Macromolecules (1980). "Primary structure" refers to the amino acid sequence of a particular peptide. "Secondary structure" refers to locally ordered, three dimensional structures within a polypeptide. These structures are commonly known as domains, e.g., enzymatic domains, extracellular domains, transmembrane domains, pore domains, and cytoplasmic tail domains. Domains are portions of a polypeptide that form a compact unit of the polypeptide and are typically 15 to 350 amino acids long. Exemplary domains include domains with enzymatic activity. Typical domains are made up of sections of lesser organization such as stretches of O-sheet and a-helieces. "Tertiary structure" refers to the complete three dimensional structure of a polypeptide monomer. "Quaternary structure" refers to the three dimensional structure formed by the noncovalent association of indepenednet tertiary units.

[0076] A "label" or a "detectable moiety" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. For example, useful labels include ³²P, fluorescent dyes, electron-dense reagents, enzymes (e.g., as commonly used in an ELISA), biotin, digoxigenin, or haptens and proteins which can be made detectable, e.g., by incorporating a radiolabel into the peptide or used to detect antibodies specifically reactive with the peptide.

[0077] The term "recombinant" when used with reference, e.g., to a cell, or nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all.

[0078] The term "heterologous" when used with reference to portions of a nucleic acid indicates that the nucleic acid comprises two or more subsequences that are not found in the same relationship to each other in nature. For instance, the nucleic acid is typically recombinantly produced, having two or more sequences from unrelated genes arranged to make a new functional nucleic acid, e.g., a promoter from one source and a coding region from another source. Similarly, a heterologous protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (e.g., a fusion protein).

[0079] The phrase "stringent hybridization conditions" refers to conditions under which a probe will hybridize to its target subsequence, typically in a complex mixture of nucleic acids, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures. An extensive guide to the hybridization of nucleic acids is found in Tijssen, Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Probes, "Overview of principles of hybridization and the strategy of nucleic acid assays" (1993). Generally, stringent conditions are selected to be about 5-10° C. lower than the thermal melting point (T_m) for the specific sequence at a defined ionic strength pH. The T_m is the temperature (under defined ionic strength, pH, and nucleic concentration) at which 50% of the probes complementary to the target hybridize to the target sequence at equilibrium (as the target sequences are present in excess, at T_m, 50% of the probes are occupied at equilibrium). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. For selective or specific hybridization, a positive signal is at least two times background, preferably 10 times background hybridization. Exemplary stringent hybridization conditions can be as following: 50% formamide, 5×SSC, and 1% SDS, incubating at 42° C., or, 5×SSC, 1% SDS, incubating at 65° C., with wash in 0.2×SSC, and 0.1% SDS at 65° C.

[0080] Nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the polypeptides which they encode are substantially identical. This occurs, for example, when a copy of a nucleic acid is created using the maximum codon degeneracy permitted by the genetic code. In such cases, the nucleic acids typically hybridize under moderately stringent hybridization conditions. Exemplary "moderately stringent hybridization conditions" include a hybridization in a buffer of 40% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 1×SSC at 45° C. A positive hybridization is at least twice background. Those of ordinary skill will readily recognize that alternative hybridization and wash conditions can be utilized to provide conditions of similar stringency. Additional guidelines for determining hybridization parameters are provided in numerous reference, e.g., and Current Protocols in Molecular Biology, ed. Ausubel, et al.

[0081] For PCR, a temperature of about 36° C. is typical for low stringency amplification, although annealing temperatures may vary between about 32° C. and 48° C. depending on primer length. For high stringency PCR amplification, a temperature of about 62° C. is typical, although high stringency annealing temperatures can range from about 50° C. to about 65° C., depending on the primer length and specificity. Typical cycle conditions for both high and low stringency amplifications include a denaturation phase of 90° C.-95° C. for 30 sec-2 min., an annealing phase lasting 30 sec.-2 min., and an extension phase of about 72° C. for 1-2 min. Protocols and guidelines for low and high stringency amplification reactions are provided, e.g., in Innis et al. (1990) PCR Protocols, A Guide to Methods and Applications, Academic Press, Inc. N.Y.).

[0082] "Antibody" refers to a polypeptide comprising a framework region from an immunoglobulin gene or fragments thereof that specifically binds and recognizes an antigen. The'recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon, and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. Typically, the antigen-binding region of an antibody will be most critical in specificity and affinity of binding.

[0083] An exemplary immunoglobulin (antibody) structural unit comprises a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (V_L) and variable heavy chain (V_H) refer to these light and heavy chains respectively.

[0084] Antibodies exist, e.g., as intact immunoglobulins or as a number of well-characterized fragments produced by digestion with various peptidases. Thus, for example, pepsin digests an antibody below the disulfide linkages in the hinge region to produce F(ab)'₂, a dimer of Fab which itself is a light chain joined to V_H-C_H1 by a disulfide bond. The F(ab)'₂ may be reduced under mild conditions to break the disulfide linkage in the hinge region, thereby converting the F(ab)'₂ dimer into an Fab' monomer. The Fab' monomer is essentially Fab with part of the hinge region (see Fundamental Immunology (Paul ed., 3d ed. 1993). While various antibody fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that such fragments may be synthesized de novo either chemically or by using recombinant DNA methodology. Thus, the term antibody, as used herein, also includes antibody fragments either produced by the modification of whole antibodies, or those synthesized de novo using recombinant DNA methodologies (e.g., single chain Fv) or those identified using phage display libraries (see, e.g., McCafferty et al., Nature 348:552-554 (1990)).

[0085] For preparation of antibodies, e.g., recombinant, monoclonal, or polyclonal antibodies, many technique known in the art can be used (see, e.g., Kohler & Milstein, Nature 256:495-497 (1975); Kozbor et al., Immunology Today 4: 72 (1983); Cole et al., pp. 77-96 in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc. (1985); Coligan, Current Protocols in Immunology (1991); Harlow & Lane, Antibodies, A Laboratory Manual (1988); and Goding, Monoclonal Antibodies: Principles and Practice (2d ed. 1986)). The genes encoding the heavy and light chains of an antibody of interest can be cloned from a cell, e.g., the genes encoding a monoclonal antibody can be cloned from a hybridoma and used to produce a recombinant monoclonal antibody. Gene libraries encoding heavy and light chains of monoclonal antibodies can also be made from hybridoma or plasma cells. Random combinations of the heavy and light chain gene products generate a large pool of antibodies with different antigenic specificity (see, e.g., Kuby, Immunology (3^rd ed. 1997)). Techniques for the production of single chain antibodies or recombinant antibodies (U.S. Pat. No. 4,946,778, U.S. Pat. No. 4,816,567) can be adapted to produce antibodies to polypeptides of this invention. Also, transgenic mice, or other organisms such as other mammals, may be used to express humanized or human antibodies (see, e.g., U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, Marks et al., Bio/Technology 10:779-783 (1992); Lonberg et al., Nature 368:856-859 (1994); Morrison, Nature 368:812-13 (1994); Fishwild et al., Nature Biotechnology 14:845-51 (1996); Neuberger, Nature Biotechnology 14:826 (1996); and Lonberg & Huszar, Intern. Rev. Immunol. 13:65-93 (1995)). Alternatively, phage display technology can be used to identify antibodies and heteromeric Fab fragments that specifically bind to selected antigens (see, e.g., McCafferty et al., Nature 348:552-554 (1990); Marks et al., Biotechnology 10:779-783 (1992)). Antibodies can also be made bispecific, i.e., able to recognize two different antigens (see, e.g., WO 93/08829, Traunecker et al., EMBO J. 10:3655-3659 (1991); and Suresh et al., Methods in Enzymology 121:210 (1986)). Antibodies can also be heteroconjugates, e.g., two covalently joined antibodies, or immunotoxins (see, e.g., U.S. Pat. No. 4,676,980, WO 91/00360; WO 92/200373; and EP 03089).

[0086] Methods for humanizing or primatizing non-human antibodies are well known in the art. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues are often referred to as import residues, which are typically taken from an import variable domain. Humanization can be essentially performed following the method of Winter and co-workers (see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-327 (1988); Verhoeyen et al., Science 239:1534-1536 (1988) and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. Accordingly, such humanized antibodies are chimeric antibodies (U.S. Pat. No. 4,816,567), wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.

[0087] A "chimeric antibody" is an antibody molecule in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species, or an entirely different molecule which confers new properties to the chimeric antibody, e.g., an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region having a different or altered antigen specificity.

[0088] The antibody can be conjugated to an "effector" moiety. The effector moiety can be any number of molecules, including labeling moieties such as radioactive labels or fluorescent labels, or can be a therapeutic moiety. In one aspect the antibody modulates the activity of the protein.

[0089] The phrase "specifically (or selectively) binds" to an antibody or "specifically (or selectively) immunoreactive with," when referring to a protein or peptide, refers to a binding reaction that is determinative of the presence of the protein, often in a heterogeneous population of proteins and other biologics. Thus, under designated immunoassay conditions, the specified antibodies bind to a particular protein at least two times the background and more typically more than 10 to 100 times background. Specific binding to an antibody under such conditions requires an antibody that is selected for its specificity for a particular protein. For example, polyclonal antibodies raised to a Bocavirus, polymorphic variants, alleles, orthologs, and conservatively modified variants, or splice variants, or portions thereof, can be selected to obtain only those polyclonal antibodies that are specifically immunoreactive with Bocavirus and not with other proteins. This selection may be achieved by subtracting out antibodies that cross-react with other molecules. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies specifically immunoreactive with a protein (see, e.g., Harlow & Lane, Antibodies, A Laboratory Manual (1988) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity).

[0090] By "therapeutically effective dose" herein is meant a dose that produces effects for which it is administered. The exact dose will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); and Pickar, Dosage Calculations (1999)).

[0091] The phrase "functional effects" in the context of assays for testing compounds that modulate activity of a Bocavirus includes the determination of a parameter that is indirectly or directly under the influence of a Bocavirus, e.g., a phenotypic or chemical effect, such as the ability to increase or decrease viral genome replication, viral RNA and protein production, virus packaging, viral particle production (particularly replication competent viral particle production), cell receptor binding, viral transduction, cellular infection, antibody binding, inducing a cellular or humoral immune response, viral protein enzymatic activity, etc. "Functional effects" include in vitro, in vivo, and ex vivo activities. Such functional effects can be measured by any means known to those skilled in the art, e.g., changes in spectroscopic characteristics (e.g., fluorescence, absorbance, refractive index); hydrodynamic (e.g., shape); chromatographic; or solubility properties for a protein; measuring inducible markers or transcriptional activation of a protein; measuring binding activity or binding assays, e.g. binding to antibodies; measuring changes in ligand or substrate binding activity; measuring viral replication; measuring cell surface marker expression; measurement of changes in protein levels; measurement of RNA stability; identification of downstream or reporter gene expression (CAT, luciferase, GFP and the like), e.g., via chemiluminescence, fluorescence, colorimetric reactions, antibody binding, and inducible markers.

[0092] "Inhibitors," "activators," and "modulators" of Bocavirus nucleic acid and polypeptide sequences are used to refer to activating, inhibitory, or modulating molecules identified using in vitro and in vivo assays of the Bocavirus nucleic acid and polypeptide sequences. Inhibitors are compounds that, e.g., bind to, partially or totally block activity, decrease, prevent, delay activation, inactivate, desensitize, or down regulate the activity or expression of Bocavirus, e.g., antagonists. "Activators" are compounds that increase, open, activate, facilitate, enhance activation, sensitize, agonize, or up regulate Bocavirus activity, e.g., agonists. Inhibitors, activators, or modulators also include genetically modified versions of Bocavirus, e.g., versions with altered activity, as well as naturally occurring and synthetic ligands, substrates, antagonists, agonists, antibodies, peptides, cyclic peptides, nucleic acids, antisense molecules, ribozymes, small chemical molecules and the like. Such assays for inhibitors and activators include, e.g., expressing Bocavirus in vitro, in cells, or cell membranes, applying putative modulator compounds, and then determining the functional effects on activity, as described above.

[0093] Samples or assays comprising Bocavirus that are treated with a potential activator, inhibitor, or modulator are compared to control samples without the inhibitor, activator, or modulator to examine the extent of inhibition. Control samples (untreated with inhibitors) are assigned a relative protein activity value of 100%. Inhibition of Bocavirus is achieved when the activity value relative to the control is about 80%, preferably 50%, more preferably 25-0%. Activation of Bocavirus is achieved when the activity value relative to the control (untreated with activators) is 110%, more preferably 150%, more preferably 200-500% (i.e., two to five fold higher relative to the control), more preferably 1000-3000% higher.

[0094] The term "test compound" or "drug candidate" or "modulator" or grammatical equivalents as used herein describes any molecule, either naturally occurring or synthetic, e.g., protein, oligopeptide (e.g., from about 5 to about 25 amino acids in length, preferably from about 10 to 20 or 12 to 18 amino acids in length, preferably 12, 15, or 18 amino acids in length), small organic molecule, polysaccharide, lipid, fatty acid, polynucleotide, oligonucleotide, etc., to be tested for the capacity to directly or indirectly modulation tumor cell proliferation. The test compound can be in the form of a library of test compounds, such as a combinatorial or randomized library that provides a sufficient range of diversity. Test compounds are optionally linked to a fusion partner, e.g., targeting compounds, rescue compounds, dimerization compounds, stabilizing compounds, addressable compounds, and other functional moieties. Conventionally, new chemical entities with useful properties are generated by identifying a test compound (called a "lead compound") with some desirable property or activity, e.g., inhibiting activity, creating variants of the lead compound, and evaluating the property and activity of those variant compounds. Often, high throughput screening (HTS) methods are employed for such an analysis.

[0095] A "small organic molecule" refers to an organic molecule, either naturally occurring or synthetic, that has a molecular weight of more than about 50 daltons and less than about 2500 daltons, preferably less than about 2000 daltons, preferably between about 100 to about 1000 daltons, more preferably between about 200 to about 500 daltons.

[0096] An "siRNA" molecule or an "RNAi" molecule refers to a nucleic acid that forms a double stranded RNA, which double stranded RNA has the ability to reduce or inhibit expression of a gene or target gene when the siRNA expressed in the same cell as the gene or target gene. "siRNA" thus refers to the double stranded RNA formed by the complementary strands. The complementary portions of the siRNA that hybridize to form the double stranded molecule typically have substantial or complete identity. In one embodiment, an siRNA refers to a nucleic acid that has substantial or complete identity to a target gene and forms a double stranded siRNA. The sequence of the siRNA can correspond to the full length target gene, or a subsequence thereof. Typically, the siRNA is at least about 15-50 nucleotides in length (e.g., each complementary sequence of the double stranded siRNA is 15-50 nucleotides in length, and the double stranded siRNA is about 15-50 base pairs in length, preferable about preferably about 20-30 base nucleotides, preferably about 20-25 or about 24-29 nucleotides in length, e.g., 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. See also PCT/US03/07237, herein incorporated by reference in its entirety.

[0097] An siRNA molecule or RNAi molecule is "specific" for a target nucleic acid if it reduces expression of the nucleic acid by at least about 10% when the siRNA or RNAi is expressed in a cell that expresses the target nucleic acid.

[0098] This invention relies on routine techniques in the field of recombinant genetics. Basic texts disclosing the general methods of use in this invention include Sambrook et al., Molecular Cloning, A Laboratory Manual (2nd ed. 1989); Kriegler, Gene Transfer and Expression: A Laboratory Manual (1990); and Current Protocols in Molecular Biology (Ausubel et al., eds., 1994)).

[0099] Bocavirus, polymorphic variants, orthologs, and alleles that are substantially identical to an amino acid sequence encoded by nucleic acids of SEQ ID NO:1 (FIG. 1) can be isolated using nucleic acid probes and oligonucleotides under stringent hybridization conditions, by screening DNA libraries or by using PCR. Genes encoding Bocavirus proteins can be isolated using cDNA libraries. Alternatively, expression libraries can be used to clone the Bocavirus, polymorphic variants, orthologs, and alleles by detecting expressed homologs immunologically with antisera or purified antibodies made against Bocavirus or portions thereof.

[0100] Other techniques that can be used to identify known and previously uncharacterized Bocavirus isolates, including representational difference analysis (RDA), DNA microarrays and use of degenerate PCR primers or other methods well known to those of skill in the art. Other methods for determining the sequence of a Bocavirus, are, for example, sequence independent single primer amplification of nucleic acids in serum (DNase-SISPA) can be used. In this method, DNA is isolated directly from environmental samples and sequenced. This method first removes contaminating human DNA in plasma or serum by DNase digestion. Viral nucleic acids protected from DNase digestion by their viral coats are then converted into double stranded DNA (dsDNA) using random primers. The dsDNA is then digested by a 4 base pair specific restriction endonuclease resulting in two overhanging bases to which are ligated a complementary oligonucleotide linker. A PCR primer complementary to the ligated linker is then used to PCR amplify the sequences between the restriction sites. The PCR products are analyzed by PAGE and distinct DNA bands are extracted, subcloned and sequenced. Similarity to known viruses is then tested using BLASTn (for nucleic acid similarity) and tBLASTx (for protein similarity). The DNase-SISPA method does not require foreknowledge of the viral sequences being amplified and can therefore theoretically amplify more divergent members of known viral families than nucleic acid sequence similarity-dependent approaches using degenerate primers or microarrays. There are several methods available and well known to those skilled in the art to obtain full-length DNAs, or extend short DNAs, for example, those based on the method of Rapid Amplification of cDNA Ends (RACE) and large scale sequencing.

[0101] To make a cDNA library to clone Bocavirus genes expressed by the genome, the source used should be rich in the RNA of choice. The mRNA is then made into cDNA using reverse transcriptase, ligated into a recombinant vector, and transfected into a recombinant host for propagation, screening and cloning. Methods for making and screening cDNA libraries are well known (see, e.g., Gubler & Hoffman, Gene 25:263-269 (1983); Sambrook et al., supra; Ausubel et al., supra).

[0102] For a genomic library, the DNA is extracted from the tissue and optionally mechanically sheared or enzymatically digested. The fragments are then separated by gradient centrifugation from undesired sizes and are constructed in suitable vectors. These vectors are packaged in vitro. Recombinant vectors can be analyzed, e.g., by plaque hybridization as described in Benton & Davis, Science 196:180-182 (1977). Colony hybridization is carried out as generally described in Grunstein et al., Proc. Natl. Acad. Sci. USA., 72:3961-3965 (1975).

[0103] A preferred method of isolating Bocavirus and orthologs, alleles, mutants, polymorphic variants, splice variants, and conservatively modified variants combines the use of synthetic oligonucleotide primers and amplification of an RNA or DNA template (see U.S. Pat. Nos. 4,683,195 and 4,683,202; PCR Protocols: A Guide to Methods and Applications (Innis et al., eds, 1990)). Methods such as polymerase chain reaction (PCR and RT-PCR) and ligase chain reaction (LCR) can be used to amplify nucleic acid sequences directly from mRNA, from cDNA, from genomic libraries or cDNA libraries. Degenerate oligonucleotides can be designed to amplify homologs using the sequences provided herein. Restriction endonuclease sites can be incorporated into the primers. Polymerase chain reaction or other in vitro amplification methods may also be useful, for example, to clone nucleic acid sequences that code for proteins to be expressed, to make nucleic acids to use as probes for detecting the presence of Bocavirus encoding mRNA in physiological samples, for nucleic acid sequencing, or for other purposes. Genes amplified by the PCR reaction can be purified from agarose gels and cloned into an appropriate vector.

[0104] Gene expression of Bocavirus can also be analyzed by techniques known in the art, e.g., reverse transcription and amplification of mRNA, isolation of total RNA or poly A.sup.+ RNA, northern blotting, dot blotting, in situ hybridization, RNase protection, high density polynucleotide array technology, e.g., and the like.

[0105] Nucleic acids encoding a Bocavirus genome or protein can be used with high density oligonucleotide array technology to identify Bocavirus, orthologs, alleles, conservatively modified variants, and polymorphic variants in this invention. In the case where the homologs being identified are linked to modulation of the cell cycle, they can be used with oligonucleotide array as a diagnostic tool in detecting the disease in a biological sample, see, e.g., Gunthand et al., AIDS Res. Hum. Retroviruses 14: 869-876 (1998); Kozal et al., Nat. Med. 2:753-759 (1996); Matson et al., Anal. Biochem. 224:110-106 (1995); Lockhart et al., Nat. Biotechnol. 14:1675-1680 (1996); Gingeras et al., Genome Res. 8:435-448 (1998); Hacia et al., Nucleic Acids Res. 26:3865-3866 (1998).

[0106] The gene of choice is typically cloned into intermediate vectors before transformation into prokaryotic or eukaryotic cells for replication and/or expression. These intermediate vectors are typically prokaryote vectors, e.g., plasmids, or shuttle vectors.

[0107] To obtain high level expression of a cloned gene or genome, one typically subclones the nucleic acid into an expression vector that contains a strong promoter to direct transcription, a transcription/translation terminator, and if for a nucleic acid encoding a protein, a ribosome binding site for translational initiation. Suitable bacterial promoters are well known in the art and described, e.g., in Sambrook et al., and Ausubel et al, supra. Bacterial expression systems for expressing the protein are available in, e.g., E. coli, Bacillus sp., and Salmonella (Palva et al., Gene 22:229-235 (1983); Mosbach et al., Nature 302:543-545 (1983). Kits for such expression systems are commercially available. Eukaryotic expression systems for mammalian cells, yeast, and insect cells are well known in the art and are also commercially available. In one preferred embodiment, retroviral expression systems are used in the present invention.

[0108] Selection of the promoter used to direct expression of a heterologous nucleic acid depends on the particular application. The promoter is preferably positioned about the same distance from the heterologous transcription start site as it is from the transcription start site in its natural setting. As is known in the art, however, some variation in this distance can be accommodated without loss of promoter function.

[0109] In addition to the promoter, the expression vector typically contains a transcription unit or expression cassette that contains all the additional elements required for the expression of the nucleic acid in host cells. A typical expression cassette thus contains a promoter operably linked to the nucleic acid sequence encoding the nucleic acid of choice and signals required for efficient polyadenylation of the transcript, ribosome binding sites, and translation termination. Additional elements of the cassette may include enhancers and, if genomic DNA is used as the structural gene, introns with functional splice donor and acceptor sites.

[0110] In addition to a promoter sequence, the expression cassette should also contain a transcription termination region downstream of the structural gene to provide for efficient termination. The termination region may be obtained from the same gene as the promoter sequence or may be obtained from different genes.

[0111] The particular expression vector used to transport the genetic information into the cell is not particularly critical. Any of the conventional vectors used for expression in eukaryotic or prokaryotic cells may be used. Standard bacterial expression vectors include plasmids such as pBR322 based plasmids, pSKF, pET23D, and fusion expression systems such as MBP, GST, and LacZ. Epitope tags can also be added to recombinant proteins to provide convenient methods of isolation, e.g., c-myc. Sequence tags may be included in an expression cassette for nucleic acid rescue. Markers such as fluorescent proteins, green or red fluorescent protein, β-gal, CAT, and the like can be included in the vectors as markers for vector transduction.

[0112] Expression vectors containing regulatory elements from eukaryotic viruses are typically used in eukaryotic expression vectors, e.g., SV40 vectors, papilloma virus vectors, retroviral vectors, and vectors derived from Epstein-Barr virus. Other exemplary eukaryotic vectors include pMSG, pAV009/A.sup.+, pMTO10/A.sup.+, pMAMneo-5, baculovirus pDSVE, and any other vector allowing expression of proteins under the direction of the CMV promoter, SV40 early promoter, SV40 later promoter, metallothionein promoter, murine mammary tumor virus promoter, Rous sarcoma virus promoter, polyhedrin promoter, or other promoters shown effective for expression in eukaryotic cells.

[0113] Expression of proteins from eukaryotic vectors can be also be regulated using inducible promoters. With inducible promoters, expression levels are tied to the concentration of inducing agents, such as tetracycline or ecdysone, by the incorporation of response elements for these agents into the promoter. Generally, high level expression is obtained from inducible promoters only in the presence of the inducing agent; basal expression levels are minimal.

[0114] In one embodiment, the vectors of the invention have a regulatable promoter, e.g., tet-regulated systems and the RU-486 system (see, e.g., Gossen & Bujard, PNAS 89:5547 (1992); Oligino et al., Gene Ther. 5:491-496 (1998); Wang et al., Gene Ther. 4:432-441 (1997); Neering et al., Blood 88:1147-1155 (1996); and Rendahl et al., Nat. Biotechnol. 16:757-761 (1998)). These impart small molecule control on the expression of the candidate target nucleic acids. This beneficial feature can be used to determine that a desired phenotype is caused by a transfected cDNA rather than a somatic mutation.

[0115] Some expression systems have markers that provide gene amplification such as thymidine kinase and dihydrofolate reductase. Alternatively, high yield expression systems not involving gene amplification are also suitable, such as using a baculovirus vector in insect cells, with a sequence of choice under the direction of the polyhedrin promoter or other strong baculovirus promoters.

[0116] The elements that are typically included in expression vectors also include a replicon that functions in E. coli, a gene encoding antibiotic resistance to permit selection of bacteria that harbor recombinant plasmids, and unique restriction sites in nonessential regions of the plasmid to allow insertion of eukaryotic sequences. The particular antibiotic resistance gene chosen is not critical, any of the many resistance genes known in the art are suitable. The prokaryotic sequences are preferably chosen such that they do not interfere with the replication of the DNA in eukaryotic cells, if necessary.

[0117] Standard transfection methods are used to produce bacterial, mammalian, yeast or insect cell lines that express large quantities of protein, which are then purified using standard techniques (see, e.g., Colley et al., J. Biol. Chem. 264:17619-17622 (1989); Guide to Protein Purification, in Methods in Enzymology, vol. 182 (Deutscher, ed., 1990)). Transformation of eukaryotic and prokaryotic cells are performed according to standard techniques (see, e.g., Morrison, J. Bact. 132:349-351 (1977); Clark-Curtiss & Curtiss, Methods in Enzymology 101:347-362 (Wu et al., eds, 1983).

[0118] Any of the well-known procedures for introducing foreign nucleotide sequences into host cells may be used. These include the use of calcium phosphate transfection, polybrene, protoplast fusion, electroporation, biolistics, liposomes, microinjection, plasma vectors, viral vectors and any of the other well known methods for introducing cloned genomic DNA, cDNA, synthetic DNA or other foreign genetic material into a host cell (see, e.g., Sambrook et al., supra). It is only necessary that the particular genetic engineering procedure used be capable of successfully introducing at least one gene into the host cell capable of expressing Bocavirus proteins and nucleic acids.

[0119] After the expression vector is introduced into the cells, the transfected cells are cultured under conditions favoring expression of the protein of choice, which is recovered from the culture using standard techniques identified below.

[0120] Either naturally occurring or recombinant Bocavirus proteins can be purified for use in diagnostic assays, for making antibodies (for diagnosis and therapy) and vaccines, and for assaying for anti-viral compounds. As described above, SEQ ID NO: 3 and SEQ ID NO:4 encode structural proteins. Naturally occurring proteins can be purified, e.g., from human tissue samples. Recombinant protein can be purified from any suitable expression system.

[0121] The protein may be purified to substantial purity by standard techniques, including selective precipitation with such substances as ammonium sulfate; column chromatography, immunopurification methods, and others (see, e.g., Scopes, Protein Purification: Principles and Practice (1982); U.S. Pat. No. 4,673,641; Ausubel et al., supra; and Sambrook et al., supra).

[0122] A number of procedures can be employed when recombinant protein is being purified. For example, proteins having established molecular adhesion properties can be reversible fused to the protein. With the appropriate ligand or substrate, a specific protein can be selectively adsorbed to a purification column and then freed from the column in a relatively pure form. The fused protein is then removed by enzymatic activity. Finally, protein could be purified using immunoaffinity columns. Recombinant protein can be purified from any suitable source, include yeast, insect, bacterial, and mammalian cells.

[0123] Methods for production and purification of recombinant protein from a bacterial or eukaryotic (e.g., yeast, mammalian cell, and the like) system are well known in the art. Recombinant proteins are expressed by transformed host cells, (e.g., bacteria) in large amounts, typically after promoter induction; but expression can be constitutive. Promoter induction with IPTG is one example of an inducible promoter system. Host cells are grown according to standard procedures in the art. Where the host cell is a bacterial cell, fresh or frozen bacteria cells are used for isolation of protein.

[0124] Recombinant proteins, particularly when expressed in bacterial host cells, may form insoluble aggregates ("inclusion bodies"). Several protocols are suitable for purification of protein inclusion bodies. For example, purification of inclusion bodies typically involves the extraction, separation and/or purification of inclusion bodies by disruption of bacterial cells, e.g., by incubation in a buffer of 50 mM TRIS/HCL pH 7.5, 50 mM NaCl, 5 mM MgCl₂, 1 mM DTT, 0.1 mM ATP, and 1 mM PMSF. The cell suspension can be lysed using 2-3 passages through a French Press, homogenized using a Polytron (Brinkman Instruments) or sonicated on ice. Alternate methods of lysing bacteria are apparent to those of skill in the art (see, e.g., Sambrook et al., supra; Ausubel et al., supra).

[0125] If necessary, the inclusion bodies are solubilized, and the lysed cell suspension is typically centrifuged to remove unwanted insoluble matter. Proteins that formed the inclusion bodies may be renatured by dilution or dialysis with a compatible buffer. Suitable solvents include, but are not limited to urea (from about 4 M to about 8 M), formamide (at least about 80%, volume/volume basis), and guanidine hydrochloride (from about 4 M to about 8 M). Some solvents which are capable of solubilizing aggregate-forming proteins, for example SDS (sodium dodecyl sulfate), 70% formic acid, are inappropriate for use in this procedure due to the possibility of irreversible denaturation of the proteins, accompanied by a lack of immunogenicity and/or activity. Although guanidine hydrochloride and similar agents are denaturants, this denaturation is not irreversible and renaturation may occur upon removal (by dialysis, for example) or dilution of the denaturant, allowing re-formation of immunologically and/or biologically active protein. Other suitable buffers are known to those skilled in the art. Human proteins are separated from other bacterial proteins by standard separation techniques, e.g., with Ni-NTA agarose resin.

[0126] Alternatively, where the host cell is a bacterium, it is possible to purify recombinant protein from bacteria periplasm. After lysis of the bacteria, the periplasmic fraction of the bacteria can be isolated by cold osmotic shock in addition to other methods known to skill in the art. To isolate recombinant proteins from the periplasm, the bacterial cells are centrifuged to form a pellet. The pellet is resuspended in a buffer containing 20% sucrose. To lyse the cells, the bacteria are centrifuged and the pellet is resuspended in ice-cold 5 mM MgSO₄ and kept in an ice bath for approximately 10 minutes. The cell suspension is centrifuged and the supernatant decanted and saved. The recombinant proteins present in the supernatant can be separated from the host proteins by standard separation techniques well known to those of skill in the art.

[0127] Standard protein separation techniques for purifying proteins are also contemplated in the present invention. Often as an initial step, particularly if the protein mixture is complex, an initial salt fractionation can separate many of the unwanted host cell proteins (or proteins derived from the cell culture media) from the recombinant protein of interest. The preferred salt is ammonium sulfate. Ammonium sulfate precipitates proteins by effectively reducing the amount of water in the protein mixture. Proteins then precipitate on the basis of their solubility. The more hydrophobic a protein is, the more likely it is to precipitate at lower ammonium sulfate concentrations. A typical protocol includes adding saturated ammonium sulfate to a protein solution so that the resultant ammonium sulfate concentration is between 20-30%. This concentration will precipitate the most hydrophobic of proteins. The precipitate is then discarded (unless the protein of interest is hydrophobic) and ammonium sulfate is added to the supernatant to a concentration known to precipitate the protein of interest. The precipitate is then solubilized in buffer and the excess salt removed if necessary, either through dialysis or diafiltration. Other methods that rely on solubility of proteins, such as cold ethanol precipitation, are well known to those of skill in the art and can be used to fractionate complex protein mixtures.

[0128] The molecular weight of the protein can be used to isolate it from proteins of greater and lesser size using ultrafiltration through membranes of different pore size (for example, Amicon or Millipore membranes). As a first step, the protein mixture is ultrafiltered through a membrane with a pore size that has a lower molecular weight cut-off than the molecular weight of the protein of interest. The retentate of the ultrafiltration is then ultrafiltered against a membrane with a molecular cut off greater than the molecular weight of the protein of interest. The recombinant protein will pass through the membrane into the filtrate. The filtrate can then be chromatographed as described below.

[0129] The protein can also be separated from other proteins on the basis of its size, net surface charge, hydrophobicity, and affinity for ligands or substrates. In addition, antibodies raised against proteins can be conjugated to column matrices and the proteins immunopurified. All of these methods are well known in the art. It will be apparent to one of skill that chromatographic techniques can be performed at any scale and using equipment from many different manufacturers (e.g., Pharmacia Biotech).

[0130] In addition to the detection of a Bocavirus gene and gene expression using nucleic acid hybridization technology, one can also use immunoassays to detect Bocavirus proteins, virus, and nucleic acids of the invention. Such assays are useful for, e.g., therapeutic and diagnostic applications. Immunoassays can be used to qualitatively or quantitatively analyze protein, virus, and nucleic acids. A general overview of the applicable technology can be found in Harlow & Lane, Antibodies: A Laboratory Manual (1988).

[0131] Methods of producing polyclonal and monoclonal antibodies that react specifically with Bocavirus protein, virus and nucleic acids are known to those of skill in the art (see, e.g., Coligan, Current Protocols in Immunology (1991); Harlow & Lane, supra; Goding, Monoclonal Antibodies: Principles and Practice (2d ed. 1986); and Kohler & Milstein, Nature 256:495-497 (1975)). Such techniques include antibody preparation by selection of antibodies from libraries of recombinant antibodies in phage or similar vectors, as well as preparation of polyclonal and monoclonal antibodies by immunizing rabbits or mice (see, e.g., Huse et al., Science 246:1275-1281 (1989); Ward et al., Nature 341:544-546 (1989)).

[0132] A number of immunogens comprising portions of a Bocavirus protein, virus or nucleic acid may be used to produce antibodies specifically reactive with the Bocavirus. For example, a recombinant Bocavirus protein or an antigenic fragment thereof, can be isolated as described herein. Recombinant protein can be expressed in eukaryotic or prokaryotic cells as described above, and purified as generally described above. Recombinant protein is the preferred immunogen for the production of monoclonal or polyclonal antibodies. Alternatively, a synthetic peptide derived from the sequences disclosed herein and conjugated to a carrier protein can be used an immunogen. Naturally occurring protein may also be used either in pure or impure form. The product is then injected into an animal capable of producing antibodies. Either monoclonal or polyclonal antibodies may be generated, for subsequent use in immunoassays to measure the protein.

[0133] Methods of production of polyclonal antibodies are known to those of skill in the art. An inbred strain of mice (e.g., BALB/C mice) or rabbits is immunized with the protein using a standard adjuvant, such as Freund's adjuvant, and a standard immunization protocol. The animal's immune response to the immunogen preparation is monitored by taking test bleeds and determining the titer of reactivity to the beta subunits. When appropriately high titers of antibody to the immunogen are obtained, blood is collected from the animal and antisera are prepared. Further fractionation of the antisera to enrich for antibodies reactive to the protein can be done if desired (see, Harlow & Lane, supra).

[0134] Monoclonal antibodies may be obtained by various techniques familiar to those skilled in the art. Briefly, spleen cells from an animal immunized with a desired antigen are immortalized, commonly by fusion with a myeloma cell (see, Kohler & Milstein, Eur. J. Immunol. 6:511-519 (1976)). Alternative methods of immortalization include transformation with Epstein Barr Virus, oncogenes, or retroviruses, or other methods well known in the art. Colonies arising from single immortalized cells are screened for production of antibodies of the desired specificity and affinity for the antigen, and yield of the monoclonal antibodies produced by such cells may be enhanced by various techniques, including injection into the peritoneal cavity of a vertebrate host. Alternatively, one may isolate DNA sequences which encode a monoclonal antibody or a binding fragment thereof by screening a DNA library from human B cells according to the general protocol outlined by Huse, et al., Science 246:1275-1281 (1989).

[0135] Monoclonal antibodies and polyclonal sera are collected and titered against the immunogen protein in an immunoassay, for example, a solid phase immunoassay with the immunogen immobilized on a solid support. Typically, polyclonal antisera with a titer of 10⁴ or greater are selected and tested for their cross reactivity against non-Bocavirus proteins and nucleic acids, using a competitive binding immunoassay. Specific polyclonal antisera and monoclonal antibodies will usually bind with a K_d of at least about 0.1 mM, more usually at least about 1 μM, preferably at least about 0.1 μM or better, and most preferably, 0.01 μM or better. Antibodies specific only for a particular Bocavirus protein can also be made by subtracting out other cross-reacting proteins, e.g., from other human Bocaviruses or other non-human Bocaviruses. In this manner, antibodies that bind only to the protein of choice may be obtained.

[0136] Once the specific antibodies against a Bocavirus protein, virus or nucleic acid in are available, the antigen can be detected by a variety of immunoassay methods. In addition, the antibody can be used therapeutically. For a review of immunological and immunoassay procedures, see Basic and Clinical Immunology (Stites & Terr eds., 7^th ed. 1991). Moreover, the immunoassays of the present invention can be performed in any of several configurations, which are reviewed extensively in Enzyme Immunoassay (Maggio, ed., 1980); and Harlow & Lane, supra.

[0137] Protein, in this case Bocavirus protein which is either associated with or separate from a Bocavirus viral particle, can be detected and/or quantified using any of a number of well recognized immunological binding assays (see, e.g., U.S. Pat. Nos. 4,366,241; 4,376,110; 4,517,288; and 4,837,168). Bocavirus viral particles may be detected based on an epitope defined by the viral proteins as presented in a viral particle and/or an epitope defined by a viral protein that is separate from a viral particle (e.g., such as may be present in an infected cell). As used in this context, then, "antigen" is meant to refer to a Bocavirus polypeptide as well as Bocavirus viral particles. For a review of the general immunoassays, see also Methods in Cell Biology: Antibodies in Cell Biology, volume 37 (Asci, ed. 1993); Basic and Clinical Immunology (Stites & Terr, eds., 7th ed. 1991). Immunological binding assays (or immunoassays) typically use an antibody that specifically binds to a protein or antigen of choice. The antibody may be produced by any of a number of means well known to those of skill in the art and as described above.

[0138] Immunoassays also often use a labeling agent to specifically bind to and label the complex formed by the antibody and antigen. The labeling agent may itself be one of the moieties comprising the antibody/antigen complex. Thus, the labeling agent may be a labeled Bocavirus protein nucleic acid or a labeled anti-Bocavirus antibody. Alternatively, the labeling agent may be a third moiety, such a secondary antibody, that specifically binds to the antibody/antigen complex (a secondary antibody is typically specific to antibodies of the species from which the first antibody is derived). Other proteins capable of specifically binding immunoglobulin constant regions, such as protein A or protein G may also be used as the label agent. These proteins exhibit a strong non-immunogenic reactivity with immunoglobulin constant regions from a variety of species (see, e.g., Kronval et al., J. Immunol. 111:1401-1406 (1973); Akerstrom et al., J. Immunol. 135:2589-2542 (1985)). The labeling agent can be modified with a detectable moiety, such as biotin, to which another molecule can specifically bind, such as streptavidin. A variety of detectable moieties are well known to those skilled in the art.

[0139] Throughout the assays, incubation and/or washing steps may be required after each combination of reagents. Incubation steps can vary from about 5 seconds to several hours, optionally from about 5 minutes to about 24 hours. However, the incubation time will depend upon the assay format, antigen, volume of solution, concentrations, and the like. Usually, the assays will be carried out at ambient temperature, although they can be conducted over a range of temperatures, such as 10° C. to 40° C.

[0140] Immunoassays for detecting Bocavirus protein, virus and nucleic acid in samples may be either competitive or noncompetitive, and may be either quantitative or non-quantitative. Noncompetitive immunoassays are assays in which antigen is directly detected and, in some instances the amount of antigen directly measured. In a "sandwich" assay, for example, the anti-Bocavirus antibodies can be bound directly to a solid substrate on which they are immobilized. These immobilized antibodies then capture the Bocavirus antigen present in the test sample. Proteins thus immobilized are then bound by a labeling agent, such as a second anti-Bocavirus antigen antibody bearing a label. Alternatively, the second antibody may lack a label, but it may, in turn, be bound by a labeled third antibody specific to antibodies of the species from which the second antibody is derived. The second or third antibody is typically modified with a detectable moiety, such as biotin, to which another molecule specifically binds, e.g., streptavidin, to provide a detectable moiety.

[0141] In competitive assays, Bocavirus antigen present in a sample is detected indirectly by detecting a decrease in a detectable signal associated with a known, added (exogenous) Bocavirus antigen displaced (competed away) from an anti-Bocavirus antigen antibody by the unknown Bocavirus antigen present in a sample. In this manner, such assays can also be adapted to provide for an indirect measurement of the amount of Bocavirus antigen present in the sample. In one competitive assay, a known amount of Bocavirus antigen is added to a sample and the sample is then contacted with an antibody that specifically binds to the Bocavirus antigen. The amount of exogenous Bocavirus antigen bound to the antibody is inversely proportional to the concentration of Bocavirus antigen present in the sample. In a particularly preferred embodiment, the antibody is immobilized on a solid substrate. The amount of Bocavirus antigen bound to the antibody may be determined either by measuring the amount of Bocavirus antigen present in Bocavirus antigen/antibody complex, or alternatively by measuring the amount of remaining uncomplexed protein. The amount of Bocavirus antigen may be detected by providing a labeled Bocavirus antigen.

[0142] A hapten inhibition assay is another competitive assay. In this assay the known Bocavirus antigen is immobilized on a solid substrate. A known amount of anti-Bocavirus antigen antibody is added to the sample, and the sample is then contacted with the immobilized Bocavirus antigen. The amount of anti-Bocavirus antigen bound to the known immobilized Bocavirus antigen is inversely proportional to the amount of Bocavirus antigen present in the sample. Again, the amount of immobilized antibody may be detected by detecting either the immobilized fraction of antibody or the fraction of the antibody that remains in solution. Detection may be direct where the antibody is labeled or indirect by the subsequent addition of a labeled moiety that specifically binds to the antibody as described above.

[0143] Immunoassays in the competitive binding format can also be used for crossreactivity determinations. For example, a Bocavirus antigen can be immobilized to a solid support. Proteins are added to the assay that compete for binding of the antisera to the immobilized antigen. The ability of the added proteins to compete for binding of the antisera to the immobilized protein is compared to the ability of the Bocavirus antigen to compete with itself. The percent crossreactivity for the above proteins is calculated, using standard calculations. Those antisera with less than 10% crossreactivity with each of the added proteins listed above are selected and pooled. The cross-reacting antibodies are optionally removed from the pooled antisera by immunoabsorption with the added considered proteins, e.g., distantly related homologs.

[0144] The immunoabsorbed and pooled antisera are then used in a competitive binding immunoassay as described above to compare a second protein, thought to be perhaps an allele or polymorphic variant of a Bocavirus antigen, to the immunogen protein. In order to make this comparison, the two proteins are each assayed at a wide range of concentrations and the amount of each protein required to inhibit 50% of the binding of the antisera to the immobilized protein is determined. If the amount of the second protein required to inhibit 50% of binding is less than 10 times the amount of the Bocavirus antigen that is required to inhibit 50% of binding, then the second protein is said to specifically bind to the polyclonal antibodies generated to Bocavirus antigen.

[0145] Western blot (immunoblot) analysis can be is used to detect and quantify the presence of Bocavirus antigen in the sample. The technique generally comprises separating sample proteins by gel electrophoresis on the basis of molecular weight, transferring the separated proteins to a suitable solid support, (such as a nitrocellulose filter, a nylon filter, or derivatized nylon filter), and incubating the sample with the antibodies that specifically bind the Bocavirus antigen. The anti-Bocavirus antigen antibodies specifically bind to the Bocavirus antigen on the solid support. These antibodies may be directly labeled or alternatively may be subsequently detected using labeled antibodies (e.g., labeled sheep anti-mouse antibodies) that specifically bind to the anti-Bocavirus antigen antibodies.

[0146] Other assay formats include liposome immunoassays (LIA), which use liposomes designed to bind specific molecules (e.g., antibodies) and release encapsulated reagents or markers. The released chemicals are then detected according to standard techniques (see Monroe et al., Amer. Clin. Prod. Rev. 5:34-41 (1986)).

[0147] One of skill in the art will appreciate that it is often desirable to minimize non-specific binding in immunoassays. Particularly, where the assay involves an antigen or antibody immobilized on a solid substrate it is desirable to minimize the amount of non-specific binding to the substrate. Means of reducing such non-specific binding are well known to those of skill in the art. Typically, this technique involves coating the substrate with a proteinaceous composition. In particular, protein compositions such as bovine serum albumin (BSA), nonfat powdered milk, and gelatin are widely used with powdered milk being most preferred.

[0148] The particular label or detectable group used in the assay is not a critical aspect of the invention, as long as it does not significantly interfere with the specific binding of the antibody used in the assay. The detectable group can be any material having a detectable physical or chemical property. Such detectable labels have been well-developed in the field of immunoassays and, in general, most any label useful in such methods can be applied to the present invention. Thus, a label is any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical or chemical means. Useful labels in the present invention include magnetic beads, fluorescent dyes (e.g., fluorescein isothiocyanate, Texas red, rhodamine, and the like), radiolabels (e.g., 3H, ¹²⁵I, ³5S, ¹⁴C, or ³²P), enzymes (e.g., horse radish peroxidase, alkaline phosphatase and others commonly used in an ELISA), and colorimetric labels such as colloidal gold or colored glass or plastic beads (e.g., polystyrene, polypropylene, latex, etc.).

[0149] The label may be coupled directly or indirectly to the desired component of the assay according to methods well known in the art. As indicated above, a wide variety of labels may be used, with the choice of label depending on sensitivity required, ease of conjugation with the compound, stability requirements, available instrumentation, and disposal provisions.

[0150] Non-radioactive labels are often attached by indirect means. Generally, a ligand molecule (e.g., biotin) is covalently bound to the molecule. The ligand then binds to another molecules (e.g., streptavidin) molecule, which is either inherently detectable or covalently bound to a signal system, such as a detectable enzyme, a fluorescent compound, or a chemiluminescent compound. The ligands and their targets can be used in any suitable combination with antibodies that recognize Bocavirus antigen, or secondary antibodies that recognize anti-Bocavirus antigen.

[0151] The molecules can also be conjugated directly to signal generating compounds, e.g., by conjugation with an enzyme or fluorophore. Enzymes of interest as labels will primarily be hydrolases, particularly phosphatases, esterases and glycosidases, or oxidotases, particularly peroxidases. Fluorescent compounds include fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, etc. Chemiluminescent compounds include luciferin, and 2,3-dihydrophthalazinediones, e.g., luminol. For a review of various labeling or signal producing systems that may be used, see U.S. Pat. No. 4,391,904.

[0152] Means of detecting labels are well known to those of skill in the art. Thus, for example, where the label is a radioactive label, means for detection include a scintillation counter or photographic film as in autoradiography. Where the label is a fluorescent label, it may be detected by exciting the fluorochrome with the appropriate wavelength of light and detecting the resulting fluorescence. The fluorescence may be detected visually, by the use of electronic detectors such as charge coupled devices (CCDs) or photomultipliers and the like. Similarly, enzymatic labels may be detected by providing the appropriate substrates for the enzyme and detecting the resulting reaction product. Colorimetric or chemiluminescent labels may be detected simply by observing the color associated with the label. Thus, in various dipstick assays, conjugated gold often appears pink, while various conjugated beads appear the color of the bead.

[0153] Some assay formats do not require the use of labeled components. For instance, agglutination assays can be used to detect the presence of the target antibodies. In this case, antigen-coated particles are agglutinated by samples comprising the target antibodies. In this format, none of the components need be labeled and the presence of the target antibody is detected by simple visual inspection.

[0154] The present invention provides diagnostic assays to detect Bocavirus, Bocavirus nucleic acids (genome and genes), Bocavirus antibodies in an infected subject, and Bocavirus proteins. In one embodiment, Bocavirus nucleic acid is detected using a nucleic acid amplification-based assay, such as a PCR assay, e.g., in a quantitative assay to determine viral load. In another embodiment, Bocavirus antigens are detected using a serological assay with antibodies (either monoclonal or polyclonal) to antigens encoded by Bocavirus.

[0155] In one embodiment of the present invention, the presence of Bocavirus, Bocavirus nucleic acid, or Bocavirus protein in a sample is determined by an immunoassay. Enzyme mediated immunoassays such as immunofluorescence assays (IFA), enzyme linked immunosorbent assays (ELISA) and immunoblotting (western) assays can be readily adapted to accomplish the detection of the Bocavirus or Bocavirus proteins. An ELISA method effective for the detection of the virus can, for example, be as follows: (1) bind an anti-Bocavirus antibody or antigen to a substrate; (2) contact the bound receptor with a fluid or tissue sample containing the virus, a viral antigen, or antibodies to the virus; (3) contact the above with an antibody bound to a detectable moiety (e.g., horseradish peroxidase enzyme or alkaline phosphatase enzyme); (4) contact the above with the substrate for the enzyme; (5) contact the above with a color reagent; (6) observe color change. The above method can be readily modified to detect presence of an anti-Bocavirus antibody in the sample or a specific Bocavirus protein as well as the virus.

[0156] Another immunologic technique that can be useful in the detection of Bocaviruses is the competitive inhibition assay, utilizing monoclonal antibodies (MABs) specifically reactive with the virus. Briefly, serum or other body fluids from the subject is reacted with an antibody bound to a substrate (e.g. an ELISA 96-well plate). Excess serum is thoroughly washed away. A labeled (enzyme-linked, fluorescent, radioactive, etc.) monoclonal antibody is then reacted with the previously reacted Bocavirus virus-antibody complex. The amount of inhibition of monoclonal antibody binding is measured relative to a control. MABs can also be used for detection directly in samples by IFA for MABs specifically reactive for the antibody-virus complex.

[0157] Alternatively, a Bocavirus antigen and/or a patient's antibodies to the virus can be detected utilizing a capture assay. Briefly, to detect antibodies to Bocavirus in a patient sample, antibodies to the patient's immunoglobulin, e.g., anti-IgG (or IgM) are bound to a solid phase substrate and used to capture the patient's immunoglobulin from serum. A Bocavirus, or reactive fragments of a Bocavirus, are then contacted with the solid phase followed by addition of a labeled antibody. The amount of patient Bocavirus specific antibody can then be quantitated by the amount of labeled antibody binding.

[0158] Additionally, a micro-agglutination test can also be used to detect the presence of Bocavirus in test samples. Briefly, latex beads are coated with an antibody and mixed with a test sample, such that Bocavirus in the tissue or body fluids that are specifically reactive with the antibody crosslink with the receptor, causing agglutination. The agglutinated antibody-virus complexes form a precipitate, visible with the naked eye or by spectrophotometer. Other assays include serologic assays, in which the relative concentrations of IgG and IgM are measured.

[0159] In the diagnostic methods described above, the sample can be taken directly from the patient or in a partially purified form. The antibody specific for a particular Bocavirus (the primary reaction) reacts by binding to the virus. Thereafter, a secondary reaction with an antibody bound to, or labeled with, a detectable moiety can be added to enhance the detection of the primary reaction. Generally, in the secondary reaction, an antibody or other ligand which is reactive, either specifically or nonspecifically with a different binding site (epitope) of the virus will be selected for its ability to react with multiple sites on the complex of antibody and virus. Thus, for example, several molecules of the antibody in the secondary reaction can react with each complex formed by the primary reaction, making the primary reaction more detectable.

[0160] The detectable moiety can allow visual detection of a precipitate or a color change, visual detection by microscopy, or automated detection by spectrometry, radiometric measurement or the like. Examples of detectable moieties include fluorescein and rhodamine (for fluorescence microscopy), horseradish peroxidase (for either light or electron microscopy and biochemical detection), biotin-streptavidin (for light or electron microscopy) and alkaline phosphatase (for biochemical detection by color change). The detection methods and moieties used can be selected, for example, from the list above or other suitable examples by the standard criteria applied to such selections (Harlow and Lane, (1988)).

[0161] As described herein, a Bocavirus infection may also, or alternatively, be detected based on the level of a Bocavirus RNA or DNA in a biological sample. Primers from Bocavirus can be used for detection of Bocavirus, diagnosis, and determination of Bocavirus viral load. Any suitable primer can be used to detect the genome, nucleic acid sub-sequence, ORF, or protein of choice, using, e.g., methods described in US 20030104009. For example, the subject nucleic acid compositions can be used as single- or double-stranded probes or primers for the detection of Bocavirus mRNA or cDNA generated from such mRNA, as obtained may be present in a biological sample (e.g., extracts of human cells). The Bocavirus polynucleotides of the invention can also be used to generate additional copies of the polynucleotides, to generate antisense oligonucleotides, and as triple-strand forming oligonucleotides. For example, two oligonucleotide primers may be employed in a polymerase chain reaction (PCR) based assay to amplify a portion of Bocavirus cDNA derived from a biological sample, wherein at least one of the oligonucleotide primers is specific for (i.e., hybridizes to) the Bocavirus polynucleotide. The amplified cDNA is then separated and detected using techniques well known in the art, such as gel electrophoresis. Similarly, oligonucleotide probes that specifically hybridize to a Bocavirus polynucleotide may be used in a hybridization assay to detect the presence of the Bocavirus polynucleotide in a biological sample. These and other uses are described in more detail below.

[0162] Nucleic acid probes specific to Bocavirus-2 can be generated using the polynucleotide sequences disclosed herein. The probes are preferably at least about 12, 15, 16, 18, 20, 22, 24, or 25 nucleotide fragments of a contiguous sequence of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof, or other polynucleotide sequence encoding a Bocavirus-2 polypeptide. Nucleic acid probes can be less than about 200, 150, 100, 75, 60, 50, 40, 30, or 25 nucleotides in length, or may be up to 2 kb, 1.5 kb, 1 kb, 0.5 kb, 0.25 kb, 0.1 kb, or 0.05 kb in length. Probes may be 5 to 40 nucleotides in length, or 8 to 35 nucleotides, or 10 to 25 nucleotides. The probes can be produced by, for example, chemical synthesis, PCR amplification, generation from longer polynucleotides using restriction enzymes, or other methods well known in the art.

[0163] The polynucleotides of the invention, particularly where used as a probe in a diagnostic assay, can be detectably labeled. Exemplary detectable labels include, but are not limited to, radiolabels, fluorochromes, (e.g. fluorescein isothiocyanate (FITC), rhodamine, Texas Red, phycoerythrin, allophycocyanin, 6-carboxyfluorescein (6-FAM), 2',7'-dimethoxy-4',5'-dichloro-6-carboxyfluorescein, 6-carboxy-X-rhodamine (ROX), 6-carboxy-2',4',7',4,7-hexachlorofluorescein (HEX), 5-carboxyfluorescein (5-FAM) or N,N,N',N¹-tetramethyl-6-carboxyrho-damine (TAMRA)), radioactive labels, (e.g. ³²P, ³5S, and ³H), and the like. The detectable label can involve two stage systems (e.g., biotin-avidin, hapten-anti-hapten antibody, and the like).

[0164] The invention also includes solid substrates, such as arrays, comprising any of the polynucleotides described herein. The polynucleotides are immobilized on the arrays using methods known in the art. An array may have one or more different polynucleotides.

[0165] Any suitable qualitative or quantitative methods known in the art for detecting specific Bocavirus nucleic acid (e.g., RNA or DNA) can be used. Bocavirus nucleic acid can be detected by, for example, in situ hybridization in tissue sections, using methods that detect single base pair differences between hybridizing nucleic acid (e.g., using the technology described in U.S. Pat. No. 5,846,717), by reverse transcriptase-PCR, or in Northern blots containing poly A.sup.+ mRNA, and other methods well known in the art. For detection of Bocavirus polynucleotides in blood or blood-derived samples, the use of methods that allow for detection of single base pair mismatches is preferred.

[0166] Using the Bocavirus nucleic acid as a basis, nucleic acid probes (e.g., including oligomers of at least about 8 nucleotides or more) can be prepared, either by excision from recombinant polynucleotides or synthetically, which probes hybridize with the Bocavirus nucleic acid, and thus are useful in detection of Bocavirus virus in a sample, and identification of infected individuals, as well as further characterization of the viral genome(s). The probes for Bocavirus polynucleotides (natural or derived) are of a length or have a sequence which allows the detection of unique viral sequences by hybridization. While about 6-8 nucleotides may be useful, longer sequences may be preferred, e.g., sequences of about 10-12 nucleotides, or about 20 nucleotides or more. Preferably, these sequences will derive from regions which lack heterogeneity among Bocavirus viral isolates.

[0167] Nucleic acid probes can be prepared using routine methods, including automated oligonucleotide synthetic methods. A complement to any unique portion of the Bocavirus genome may be used, e.g., a portion of the Bocavirus genome that allows for distinguishing Bocavirus from other viruses that may be present in the sample. For use as probes, complete complementarity is desirable, though it may be unnecessary as the length of the fragment is increased.

[0168] For use of such probes as diagnostics, the biological sample to be analyzed, such as blood or serum, may be treated, if desired, to extract the nucleic acids contained therein. The resulting nucleic acid from the sample may be subjected to gel electrophoresis or other size separation techniques; alternatively, the nucleic acid sample may be dot blotted without size separation. The probes are usually labeled with a detectable label. Suitable labels, and methods for labeling probes are known in the art, and include, for example, radioactive labels incorporated by nick translation or kinasing, biotin, fluorescent probes, and chemiluminescent probes. The nucleic acids extracted from the sample are then treated with the labeled probe under hybridization conditions of suitable stringencies.

[0169] The probes can be made completely complementary to the Bocavirus genome or portion thereof. Therefore, usually high stringency conditions are desirable in order to prevent or at least minimize false positives. However, conditions of high stringency should only be used if the probes are complementary to regions of the viral genome which lack heterogeneity among Bocavirus viral isolates. The stringency of hybridization is determined by a number of factors during hybridization and during the washing procedure, including temperature, ionic strength, length of time, and concentration of formamide. These factors are outlined in, for example, Sambrook et al. (1989), "Molecular Cloning; A Laboratory Manual", Second Edition (Cold Spring Harbor Press, Cold Spring Harbor, N.Y.).

[0170] Generally, it is expected that the Bocavirus sequences will be present in a biological sample (e.g., blood, cells, and the liked) obtained from an infected individual at relatively low levels, e.g., at approximately 10²-10⁴ Bocavirus sequences per 10⁶ cells. This level may require that amplification techniques be used in hybridization assays. Such techniques are known in the art.

[0171] For example, the Enzo Biochemical Corporation "Bio-Bridge" system uses terminal deoxynucleotide transferase to add unmodified 3'-poly-dT-tails to a DNA probe. The poly dT-tailed probe is hybridized to the target nucleotide sequence, and then to a biotin-modified poly-A. PCT Publication No. WO84/03520 and European application No. EPAl24221 describe a DNA hybridization assay in which: (1) analyte is annealed to a single-stranded DNA probe that is complementary to an enzyme-labeled oligonucleotide; and (2) the resulting tailed duplex is hybridized to an enzyme-labeled oligonucleotide. EPA 204510 describes a DNA hybridization assay in which analyte DNA is contacted with a probe that has a tail, such as a poly-dT tail, an amplifier strand that has a sequence that hybridizes to the tail of the probe, such as a poly-A sequence, and which is capable of binding a plurality of labeled strands.

[0172] Non-PCR-based, sequence specific DNA amplification techniques can also be used in the invention to detect Bocavirus sequences. An example of such techniques include, but are not necessarily limited to the Invader assay, see, e.g., Kwiatkowski et al. Mol. Diagn. December 1999; 4(4):353-64. See also U.S. Pat. No. 5,846,717.

[0173] A particularly desirable technique may first involve amplification of the target Bocavirus sequences in sera approximately 10,000 fold, e.g., to approximately 10 sequences/mL. This may be accomplished, for example, by the polymerase chain reactions (PCR) technique described which is by Saiki et al. (1986), by Mullis, U.S. Pat. No. 4,683,195, and by Mullis et al. U.S. Pat. No. 4,683,202. Other amplification methods are well known in the art. In a preferred embodiment, a sample suspected of comprising the Bocavirus nucleic acid is contacted with at least one primer that hybridizes to a nucleotide sequence of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof, said contacting being under conditions suitable for amplification of an amplification product from a Bocavirus nucleic acid in the sample.

[0174] The probes, or alternatively nucleic acid from the samples, may be provided in solution for such assays, or may be affixed to a support (e.g., solid or semi-solid support). Examples of supports that can be used are nitrocellulose (e.g., in membrane or microtiter well form), polyvinyl chloride (e.g., in sheets or microtiter wells), polystyrene latex (e.g., in beads or microtiter plates, polyvinylidine fluoride, diazotized paper, nylon membranes, activated beads, and Protein A beads.

[0175] In one embodiment, the probe (or sample nucleic acid) is provided on an array for detection. Arrays can be created by, for example, spotting polynucleotide probes onto a substrate (e.g., glass, nitrocellulose, and the like) in a two-dimensional matrix or array. The probes can be bound to the substrate by either covalent bonds or by non-specific interactions, such as hydrophobic interactions. Samples of polynucleotides can be detectably labeled (e.g., using radioactive or fluorescent labels) and then hybridized to the probes. Double stranded polynucleotides, comprising the labeled sample polynucleotides bound to probe polynucleotides, can be detected once the unbound portion of the sample is washed away. Techniques for constructing arrays and methods of using these arrays are described in EP 799 897; WO 97/29212; WO 97/27317; EP 785 280; WO 97/02357; U.S. Pat. No. 5,593,839; U.S. Pat. No. 5,578,832; EP 728 520; U.S. Pat. No. 5,599,695; EP 721 016; U.S. Pat. No. 5,556,752; WO 95/22058; and U.S. Pat. No. 5,631,734. Arrays are particularly useful where, for example a single sample is to be analyzed for the presence of two or more nucleic acid target regions, as the probes for each of the target regions, as well as controls (both positive and negative) can be provided on a single array. Arrays thus facilitate rapid and convenience analysis.

[0176] The invention further provides diagnostic reagents and kits comprising one or more such reagents for use in a variety of diagnostic assays, including for example, immunoassays such as ELISA and "sandwich"-type immunoassays, as well as nucleic acid assay, e.g., PCR assays. In a related embodiment, the assay is performed in a flow-through or strip test format, wherein the binding agent is immobilized on a membrane, such as nitrocellulose. Such kits may preferably include at least a first peptide, or a first antibody or antigen binding fragment of the invention, a functional fragment thereof, or a cocktail thereof, or a first oligo pair, and means for signal generation. The kit's components may be pre-attached to a solid support, or may be applied to the surface of a solid support when the kit is used. The signal generating means may come pre-associated with an antibody or nucleic acid of the invention or may require combination with one or more components, e.g., buffers, nucleic acids, antibody-enzyme conjugates, enzyme substrates, or the like, prior to use.

[0177] Kits may also include additional reagents, e.g., blocking reagents for reducing nonspecific binding to the solid phase surface, washing reagents, enzyme substrates, enzymes, and the like. The solid phase surface may be in the form of microtiter plates, microspheres, or other materials suitable for immobilizing nucleic acids, proteins, peptides, or polypeptides. An enzyme that catalyzes the formation of a chemiluminescent or chromogenic product or the reduction of a chemiluminescent or chromogenic substrate is one such component of the signal generating means. Such enzymes are well known in the art. Where a radiolabel, chromogenic, fluorogenic, or other type of detectable label or detecting means is included within the kit, the labeling agent may be provided either in the same container as the diagnostic or therapeutic composition itself, or may alternatively be placed in a second distinct container means into which this second composition may be placed and suitably aliquoted. Alternatively, the detection reagent and the label may be prepared in a single container means, and in most cases, the kit will also typically include a means for containing the vial(s) in close confinement for commercial sale and/or convenient packaging and delivery.

[0178] Assays for modulators of Bocavirus are also contemplated in the present invention. Modulation of a Bocavirus, and corresponding modulation of the cell cycle, e.g., tumor cell, proliferation, can be assessed using a variety of in vitro and in vivo assays, including cell-based models. Such assays can be used to test for inhibitors and activators of Bocavirus. Modulators of Bocavirus are tested using either recombinant or naturally occurring protein of choice, preferably human Bocavirus.

[0179] Preferably, the Bocavirus will have the sequence as shown in SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, or SEQ ID NO:49. Alternatively, the Bocavirus of the assay will be derived from a eukaryote and encode an amino acid subsequence having substantial amino acid sequence identity to a sequence as shown in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, or SEQ ID NO:54. Generally, the amino acid sequence identity will be at least 60%, preferably at least 65%, 70%, 75%, 80%, 85%, or 90%, most preferably at least 95%.

[0180] Measurement of modulation of a Bocavirus or a cell expressing Bocavirus, either recombinant or naturally occurring, can be performed using a variety of assays, in vitro, in vivo, and ex vivo, as described herein. A suitable physical, chemical or phenotypic change that affects activity, e.g., enzymatic activity, cell surface marker expression, viral replication and proliferation can be used to assess the influence of a test compound on the polypeptide of this invention. When the functional effects are determined using intact cells or animals, one can also measure a variety of effects.

[0181] Assays to identify compounds with Bocavirus modulating activity can be performed in vitro. Such assays can used full length Bocavirus or a variant thereof, or a mutant thereof, or a fragment thereof, such as a RING domain. Purified recombinant or naturally occurring protein can be used in the in vitro methods of the invention. In addition to purified Bocavirus, the recombinant or naturally occurring protein can be part of a cellular lysate or a cell membrane. As described below, the binding assay can be either solid state or soluble. Preferably, the protein or membrane is bound to a solid support, either covalently or non-covalently. Often, the in vitro assays of the invention are substrate or ligand binding or affinity assays, either non-competitive or competitive. Other in vitro assays include measuring changes in spectroscopic (e.g., fluorescence, absorbance, refractive index), hydrodynamic (e.g., shape), chromatographic, or solubility properties for the protein.

[0182] In one embodiment, a high throughput binding assay is performed in which the protein or a fragment thereof is contacted with a potential modulator and incubated for a suitable amount of time. In one embodiment, the potential modulator is bound to a solid support, and the protein is added. In another embodiment, the protein is bound to a solid support. A wide variety of modulators can be used, as described below, including small organic molecules, peptides, antibodies, etc. A wide variety of assays can be used to identify Bocavirus-modulator binding, including labeled protein-protein binding assays, electrophoretic mobility shifts, immunoassays, enzymatic assays, and the like. In some cases, the binding of the candidate modulator is determined through the use of competitive binding assays, where interference with binding of a known ligand or substrate is measured in the presence of a potential modulator. Either the modulator or the known ligand or substrate is bound first, and then the competitor is added. After the protein is washed, interference with binding, either of the potential modulator or of the known ligand or substrate, is determined. Often, either the potential modulator or the known ligand or substrate is labeled.

[0183] In another embodiment, the Bocavirus is expressed in a cell, and functional, e.g., physical and chemical or phenotypic, changes are assayed to identify modulators of the cell cycle. Any suitable functional effect can be measured, as described herein. The Bocavirus can be naturally occurring or recombinant. Also, fragments of the Bocavirus or chimeric proteins can be used in cell based assays. In addition, point mutants in essential residues required by the catalytic site can be used in these assays.

[0184] The compounds tested as modulators of Bocavirus can be any small organic molecule, or a biological entity, such as a protein, e.g., an antibody or peptide, a sugar, a nucleic acid, e.g., an antisense oligonucleotide or a ribozyme or RNAi, or a lipid. Alternatively, modulators can be genetically altered versions of a Bocavirus. Typically, test compounds will be small organic molecules, peptides, circular peptides, RNAi, antisense molecules, ribozymes, and lipids.

[0185] Essentially any chemical compound can be used as a potential modulator or ligand in the assays of the invention, although most often compounds can be dissolved in aqueous or organic (especially DMSO-based) solutions are used. The assays are designed to screen large chemical libraries by automating the assay steps and providing compounds from any convenient source to assays, which are typically run in parallel (e.g., in microtiter formats on microtiter plates in robotic assays). It will be appreciated that there are many suppliers of chemical compounds, including Sigma (St. Louis, Mo.), Aldrich (St. Louis, Mo.), Sigma-Aldrich (St. Louis, Mo.), Fluka Chemika-Biochemica Analytika (Buchs Switzerland) and the like.

[0186] In one preferred embodiment, high throughput screening methods involve providing a combinatorial small organic molecule or peptide library containing a large number of potential therapeutic compounds (potential modulator or ligand compounds). Such "combinatorial chemical libraries" or "ligand libraries" are then screened in one or more assays, as described herein, to identify those library members (particular chemical species or subclasses) that display a desired characteristic activity. The compounds thus identified can serve as conventional "lead compounds" or can themselves be used as potential or actual therapeutics.

[0187] A combinatorial chemical library is a collection of diverse chemical compounds generated by either chemical synthesis or biological synthesis, by combining a number of chemical "building blocks" such as reagents. For example, a linear combinatorial chemical library such as a polypeptide library is formed by combining a set of chemical building blocks (amino acids) in every possible way for a given compound length (i.e., the number of amino acids in a polypeptide compound). Millions of chemical compounds can be synthesized through such combinatorial mixing of chemical building blocks.

[0188] Preparation and screening of combinatorial chemical libraries is well known to those of skill in the art. Such combinatorial chemical libraries include, but are not limited to, peptide libraries (see, e.g., U.S. Pat. No. 5,010,175, Furka, Int. J. Pept. Prot. Res. 37:487-493 (1991) and Houghton et al., Nature 354:84-88 (1991)). Other chemistries for generating chemical diversity libraries can also be used. Such chemistries include, but are not limited to: peptoids (e.g., PCT Publication No. WO 91/19735), encoded peptides (e.g., PCT Publication No. WO 93/20242), random bio-oligomers (e.g., PCT Publication No. WO 92/00091), benzodiazepines (e.g., U.S. Pat. No. 5,288,514), diversomers such as hydantoins, benzodiazepines and dipeptides (Hobbs et al., Proc. Nat. Acad. Sci. USA 90:6909-6913 (1993)), vinylogous polypeptides (Hagihara et al., J. Amer. Chem. Soc. 114:6568 (1992)), nonpeptidal peptidomimetics with glucose scaffolding (Hirschmann et al., J. Amer. Chem. Soc. 114:9217-9218 (1992)), analogous organic syntheses of small compound libraries (Chen et al., J. Amer. Chem. Soc. 116:2661 (1994)), oligocarbamates (Cho et al., Science 261:1303 (1993)), and/or peptidyl phosphonates (Campbell et al., J. Org. Chem. 59:658 (1994)), nucleic acid libraries (see Ausubel, Berger and Sambrook, all supra), peptide nucleic acid libraries (see, e.g., U.S. Pat. No. 5,539,083), antibody libraries (see, e.g., Vaughn et al., Nature Biotechnology, 14(3):309-314 (1996) and PCT/US96/10287), carbohydrate libraries (see, e.g., Liang et al., Science, 274:1520-1522 (1996) and U.S. Pat. No. 5,593,853), small organic molecule libraries (see, e.g., benzodiazepines, Baum C&EN, January 18, page 33 (1993); isoprenoids, U.S. Pat. No. 5,569,588; thiazolidinones and metathiazanones, U.S. Pat. No. 5,549,974; pyrrolidines, U.S. Pat. Nos. 5,525,735 and 5,519,134; morpholino compounds, U.S. Pat. Nos. 5,506,337; benzodiazepines, 5,288,514, and the like).

[0189] Devices for the preparation of combinatorial libraries are commercially available (see, e.g., 357 MPS, 390 MPS, Advanced Chem Tech, Louisville Ky., Symphony, Rainin, Woburn, Mass., 433A Applied Biosystems, Foster City, Calif., 9050 Plus, Millipore, Bedford, Mass.). In addition, numerous combinatorial libraries are themselves commercially available (see, e.g., ComGenex, Princeton, N.J., Asinex, Moscow, Ru, Tripos, Inc., St. Louis, Mo., ChemStar, Ltd, Moscow, RU, 3D Pharmaceuticals, Exton, Pa., Martek Biosciences, Columbia, Md., etc.).

[0190] In one embodiment the invention, soluble assays using a Bocavirus, or a cell or tissue expressing an Bocavirus, either naturally occurring or recombinant are provided. In another embodiment, the invention provides solid phase based in vitro assays in a high throughput format, where the Bocavirus is attached to a solid phase. Any one of the assays described herein can be adapted for high throughput screening.

[0191] In the high throughput assays of the invention, either soluble or solid state, it is possible to screen up to several thousand different modulators or ligands in a single day. This methodology can be used for Bocavirus in vitro, or for cell-based or membrane-based assays comprising a Bocavirus. In particular, each well of a microtiter plate can be used to run a separate assay against a selected potential modulator, or, if concentration or incubation time effects are to be observed, every 5-10 wells can test a single modulator. Thus, a single standard microtiter plate can assay about 100 (e.g., 96) modulators. If 1536 well plates are used, then a single plate can easily assay from about 100- about 1500 different compounds. It is possible to assay many plates per day; assay screens for up to about 6,000, 20,000, 50,000, or more than 100,000 different compounds are possible using the integrated systems of the invention.

[0192] For a solid state reaction, the protein of interest or a fragment thereof, e.g., an extracellular domain, or a cell or membrane comprising the protein of interest or a fragment thereof as part of a fusion protein can be bound to the solid state component, directly or indirectly, via covalent or non covalent linkage. A tag for covalent or non-covalent binding can be any of a variety of components. In general, a molecule which binds the tag (a tag binder) is fixed to a solid support, and the tagged molecule of interest is attached to the solid support by interaction of the tag and the tag binder.

[0193] A number of tags and tag binders can be used, based upon known molecular interactions well described in the literature. For example, where a tag has a natural binder, for example, biotin, protein A, or protein G, it can be used in conjunction with appropriate tag binders (avidin, streptavidin, neutravidin, the Fc region of an immunoglobulin, etc.) Antibodies to molecules with natural binders such as biotin are also widely available and appropriate tag binders; see, SIGMA Immunochemicals 1998 catalogue SIGMA, St. Louis Mo.).

[0194] Similarly, any haptenic or antigenic compound can be used in combination with an appropriate antibody to form a tag/tag binder pair. Thousands of specific antibodies are commercially available and many additional antibodies are described in the literature. For example, in one common configuration, the tag is a first antibody and the tag binder is a second antibody which recognizes the first antibody. In addition to antibody-antigen interactions, receptor-ligand interactions are also appropriate as tag and tag-binder pairs. For example, agonists and antagonists of cell membrane receptors (e.g., cell receptor-ligand interactions such as transferrin, c-kit, viral receptor ligands, cytokine receptors, chemokine receptors, interleukin receptors, immunoglobulin receptors and antibodies, the cadherein family, the integrin family, the selectin family, and the like; see, e.g., Pigott & Power, The Adhesion Molecule Facts Book I (1993). Similarly, toxins and venoms, viral epitopes, hormones (e.g., opiates, steroids, etc), intracellular receptors (e.g. which mediate the effects of various small ligands, including steroids, thyroid hormone, retinoids and vitamin D; peptides), drugs, lectins, sugars, nucleic acids (both linear and cyclic polymer configurations), oligosaccharides, proteins, phospholipids and antibodies can all interact with various cell receptors.

[0195] Synthetic polymers, such as polyurethanes, polyesters, polycarbonates, polyureas, polyamides, polyethyleneimines, polyarylene sulfides, polysiloxanes, polyimides, and polyacetates can also form an appropriate tag or tag binder. Many other tag/tag binder pairs are also useful in assay systems described herein, as would be apparent to one of skill upon review of this disclosure.

[0196] Common linkers such as peptides, polyethers, and the like can also serve as tags, and include polypeptide sequences, such as poly gly sequences of between about 5 and 200 amino acids. Such flexible linkers are known to persons of skill in the art. For example, poly(ethelyne glycol) linkers are available from Shearwater Polymers, Inc. Huntsville, Ala. These linkers optionally have amide linkages, sulfhydryl linkages, or heterofunctional linkages.

[0197] Tag binders are fixed to solid substrates using any of a variety of methods currently available. Solid substrates are commonly derivatized or functionalized by exposing all or a portion of the substrate to a chemical reagent which fixes a chemical group to the surface which is reactive with a portion of the tag binder. For example, groups which are suitable for attachment to a longer chain portion would include amines, hydroxyl, thiol, and carboxyl groups. Aminoalkylsilanes and hydroxyalkylsilanes can be used to functionalize a variety of surfaces, such as glass surfaces. The construction of such solid phase biopolymer arrays is well described in the literature. See, e.g., Merrifield, J. Am. Chem. Soc. 85:2149-2154 (1963) (describing solid phase synthesis of, e.g., peptides); Geysen et al., J. Immun. Meth. 102:259-274 (1987) (describing synthesis of solid phase components on pins); Frank & Doring, Tetrahedron 44:60316040 (1988) (describing synthesis of various peptide sequences on cellulose disks); Fodor et al., Science, 251:767-777 (1991); Sheldon et al., Clinical Chemistry 39(4):718-719 (1993); and Kozal et al., Nature Medicine 2(7):753759 (1996) (all describing arrays of biopolymers fixed to solid substrates). Non-chemical approaches for fixing tag binders to substrates include other common methods, such as heat, cross-linking by UV radiation, and the like.

[0198] Within certain aspects, Bocavirus, proteins or peptides and immunogenic fragments thereof, and/or polynucleotides, as well as anti-Bocavirus antibodies and/or T cells, may be incorporated into pharmaceutical compositions or immunogenic compositions (e.g., vaccines). Whole virus vaccine (live and attenuated, or replication incompetent, or killed) or subunit vaccines, such as structural or non-structural Bocavirus proteins or immunogenic fragments thereof, of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54 or conservatively modified variants thereof, can be used to treat or prevent Bocavirus infections by eliciting an immune response in a subject. Alternatively, a pharmaceutical composition may comprise an antigen-presenting cell (e.g., a dendritic cell) transfected with a Bocavirus polynucleotide such that the antigen-presenting cell expresses a Bocavirus peptide.

[0199] Pharmaceutical compositions comprise one or more such vaccine compounds and a physiologically acceptable carrier. Vaccines may comprise one or more such compounds and a non-specific immune response enhancer. A non-specific immune response enhancer may be any substance that enhances an immune response to an exogenous antigen. Examples of non-specific immune response enhancers include adjuvants, biodegradable microspheres (e.g., polylactic galactide) and liposomes (into which the compound is incorporated; see, e.g., U.S. Pat. No. 4,235,877). Most adjuvants contain a substance designed to protect the antigen from rapid catabolism, such as aluminum hydroxide or mineral oil, and a stimulator of immune responses, such as lipid A, Bortadella pertussis or Mycobacterium tuberculosis derived proteins. Suitable adjuvants are commercially available as, for example, Freund's Incomplete Adjuvant and Complete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.); AS-2 (SmithKline Beecham); aluminum salts such as aluminum hydroxide gel (alum) or aluminum phosphate; salts of calcium, iron or zinc; an insoluble suspension of acylated tyrosine; acylated sugars; cationically or anionically derivatized polysaccharides; polyphosphazenes; biodegradable microspheres; monophosphoryl lipid A and quil A. Cytokines, such as GM-CSF or interleukin-2, -7, or -12, may also be used as adjuvants.

[0200] Vaccine preparation is generally described in, for example, Powell and Newman, eds., Vaccine Design (the subunit and adjuvant approach), Plenum Press (NY, 1995). Vaccines may be designed to generate antibody immunity and/or cellular immunity such as that arising from CTL or CD4+ T cells.

[0201] Pharmaceutical compositions and vaccines within the scope of the present invention may also contain other compounds, which may be biologically active or inactive. For example, one or more immunogenic portions of other antigens may be present, either incorporated into a fusion polypeptide or as a separate compound, within the composition or vaccine. Polypeptides may, but need not, be conjugated to other macromolecules as described, for example, within U.S. Pat. Nos. 4,372,945 and 4,474,757. Pharmaceutical compositions and vaccines may generally be used for prophylactic and therapeutic purposes.

[0202] Nucleic acid vaccines encoding a genome, structural protein or non-structural protein or a fragment thereof of Bocavirus can also be used to elicit an immune response to treat or prevent Bocavirus infection. Numerous gene delivery techniques are well known in the art, such as those described by Rolland (1998) Crit. Rev. Therap. Drug Carrier Systems 15:143-198, and references cited therein. Appropriate nucleic acid expression systems contain the necessary DNA sequences for expression in the patient (such as a suitable promoter and terminating signal). In a preferred embodiment, the DNA may be introduced using a viral expression system (e.g., vaccinia, pox virus, retrovirus, or adenovirus), which may involve the use of a non-pathogenic (defective), replication competent virus. Suitable systems are disclosed, for example, in Fisher-Hoch et al. (1989) Proc. Natl. Acad. Sci. USA 86:317-321; Flexner et al. (1989) Ann. N.Y. Acad. Sci. 569:86-103; Flexner et al. (1990) Vaccine 8:17-21; U.S. Pat. Nos. 4,603,112, 4,769,330, 4,777,127 and 5,017,487; WO 89/01973; GB 2,200,651; EP 0,345,242; WO 91/02805; Berkner (1988) Biotechniques 6:616-627; Rosenfeld et al. (1991) Science 252:431-434; Kolls et al. (1994) Proc. Natl. Acad. Sci. USA 91:215-219; Kass-Eisler et al. (1993) Proc. Natl. Acad. Sci. USA 90:11498-11502; Guzman et al. (1993) Circulation 88:2838-2848; and Guzman et al. (1993) Cir. Res. 73:1202-1207. Techniques for incorporating DNA into such expression systems are well known to those of ordinary skill in the art. The DNA may also be "naked," as described, for example, in Ulmer et al. (1993) Science 259:1745-1749 and reviewed by Cohen (1993) Science 259:1691-1692. The uptake of naked DNA may be increased by coating the DNA onto biodegradable beads, which are efficiently transported into the cells. It will be apparent that a vaccine may comprise both a polynucleotide and a polypeptide component. Such vaccines may provide for an enhanced immune response.

[0203] Vaccines and pharmaceutical compositions may be presented in unit-dose or multi-dose containers, such as sealed ampoules or vials. Such containers are preferably hermetically sealed to preserve sterility of the formulation until use. In general, formulations may be stored as suspensions, solutions or emulsions in oily or aqueous vehicles. Alternatively, a vaccine or pharmaceutical composition may be stored in a freeze-dried condition requiring only the addition of a sterile liquid carrier immediately prior to use.

[0204] Pharmaceutically acceptable carriers are determined in part by the particular composition being administered (e.g., nucleic acid, protein, modulatory compounds or transduced cell), as well as by the particular method used to administer the composition. Accordingly, there are a wide variety of suitable formulations of pharmaceutical compositions of the present invention (see, e.g., Remington's Pharmaceutical Sciences, 17^th ed., 1989). Administration can be in any convenient manner, e.g., by injection, oral administration, inhalation, transdermal application, or rectal administration.

[0205] Formulations suitable for oral administration can consist of (a) liquid solutions, such as an effective amount of the packaged nucleic acid suspended in diluents, such as water, saline or PEG 400; (b) capsules, sachets or tablets, each containing a predetermined amount of the active ingredient, as liquids, solids, granules or gelatin; (c) suspensions in an appropriate liquid; and (d) suitable emulsions. Tablet forms can include one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphates, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid, and other excipients, colorants, fillers, binders, diluents, buffering agents, moistening agents, preservatives, flavoring agents, dyes, disintegrating agents, and pharmaceutically compatible carriers. Lozenge forms can comprise the active ingredient in a flavor, e.g., sucrose, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin or sucrose and acacia emulsions, gels, and the like containing, in addition to the active ingredient, carriers known in the art.

[0206] The compound of choice, alone or in combination with other suitable components, can be made into aerosol formulations (i.e., they can be "nebulized") to be administered via inhalation. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like.

[0207] Formulations suitable for parenteral administration, such as, for example, by intraarticular (in the joints), intravenous, intramuscular, intradermal, intraperitoneal, and subcutaneous routes, include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. In the practice of this invention, compositions can be administered, for example, by intravenous infusion, orally, topically, intraperitoneally, intravesically or intrathecally. Parenteral administration and intravenous administration are the preferred methods of administration. The formulations of commends can be presented in unit-dose or multi-dose sealed containers, such as ampules and vials.

[0208] Such compositions may also comprise buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants, bacteriostats, chelating agents such as EDTA or glutathione, adjuvants (e.g., aluminum hydroxide), solutes that render the formulation isotonic, hypotonic or weakly hypertonic with the blood of a recipient, suspending agents, thickening agents and/or preservatives. Alternatively, compositions of the present invention may be formulated as a lyophilizate. Compounds may also be encapsulated within liposomes using well known technology.

[0209] Injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described. Cells transduced by nucleic acids for ex vivo therapy can also be administered intravenously or parenterally as described above.

[0210] The dose administered to a patient, in the context of the present invention should be sufficient to effect a beneficial therapeutic response in the patient over time. The dose will be determined by the efficacy of the particular vector employed and the condition of the patient, as well as the body weight or surface area of the patient to be treated. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects that accompany the administration of a particular vector, or transduced cell type in a particular patient.

[0211] In determining the effective amount of the vector to be administered in the treatment or prophylaxis of conditions owing to diminished or aberrant expression of the protein, the physician evaluates circulating plasma levels of the vector, vector toxicities, progression of the disease, and the production of anti-vector antibodies. In general, the dose equivalent of a naked nucleic acid from a vector is from about 1 μg to 100 μg for a typical 70 kilogram patient, and doses of vectors are calculated to yield an equivalent amount of therapeutic nucleic acid.

[0212] For administration, compounds and transduced cells of the present invention can be administered at a rate determined by the LD-50 of the inhibitor, vector, or transduced cell type, and the side-effects of the inhibitor, vector or cell type at various concentrations, as applied to the mass and overall health of the patient. Administration can be accomplished via single or divided doses.

Example

[0213] Using viral metagenomics a novel parvovirus species (termed HBoV2) and variants thereof were identified in human stool, whose closest phylogenetic relative is the human bocavirus (HBoV). HBoV2 has an identical genomic organization to HBoV but shares only 78%, 67%, and 80% identity to its NS1, NP1 and VP1/VP2 proteins. Using PCR HBoV2 sequences were detected in 5/98 Pakistani children stool samples and 3/699 stool samples from the UK. Near full genome sequencing showed the presence of three divergent genotypes and evidence of recombination.

[0214] Patients, materials, and methods. Stool samples from 57 children with non-polio AFP (mean age 54.6 months) and 41 from healthy Pakistani children (mean age 39.8 months) were analyzed. Samples were collected as part of the WHO poliovirus eradication program. 699 stool samples from a mixed age population submitted for enteric bacteriology screening in Edinburgh were also analyzed.

[0215] Stool supernatants were processed for viral metagenomics as previously described (Kapoor et al., J Virol 82:311-20, 2008) with minor modifications. Briefly, stool samples were suspended in Hanks buffered salt solution (HBSS), centrifuged at 12,000 rpm for 2 minutes in a tabletop microfuge and the supernatant collected. The supernatant was then filtered (Ultrafree-MC HV 0.45 um sterile filter, Millipore, UFC30HV0S). The flow through was then ultracentrifuged at 30,000 rpm for 3 hrs at 10° C. (Beckman Coulter Optima LE-80 ultracentrifuge) to pellet viral particles (and other particles), which were then re-suspended in 100 μl HBBS which was then treated with 14 U of Turbo DNase (Ambion) and 2 μl of 10 mg/ml RNase A at 37 C for 2 hours to digest non-particle protected (naked) nucleic acids. 140 μl total viral nucleic acids (both RNA and DNA) were extracted with QIAAMP Viral RNA Mini Kit and released into 40 μl water plus 20 U recombinant RNase inhibitor (Roche). 11 μl of extracted nucleic acid was treated for viral RNA amplification as described (Kapoor et al., J Virol 82:311-20, 2008). 11 μl was also mixed with 1.5 μl of 10×Ecopol buffer (New England Biolabs), a 1 μl solution containing each dNTP at 10 mM, and 1 μl of 50 μM primer RA01 (GCCGGAGCTCTGCAGATATC) (SEQ ID NO:60). After a denaturation step at 94° C. for 3 min and chilling on ice (to anneal primer RA01 to DNA), 2.5 units of 3'-5' exo-Klenow DNA polymerase (New England Biolabs) was added to extend RA01 and incubated at 37° C. for 1 h. The denaturation, RA01 annealing, Klenow elongation steps was then repeated once with the addition of 2.5 units of fresh 3'-5' exo Klenow DNA polymerase after chilling followed by 75° C. for 10 min. 7.5 μl of the Klenow-treated product was then used as a template in a subsequent 50 ul PCR consisting of AMPLITAQ GOLD PCR buffer II (10 mM Tris-HCl [pH 8.3], 50 mM KCl) (Applied Biosystems), 3 mM MgCl₂, each dNTP at 0.3 mM, 1 μM of primer RA02 (GCCGGAGCTCTGCAGATATC) (SEQ ID NO:61), and 2.5 units of AMPLITAQ GOLD DNA polymerase LD (Applied Biosystems). An initial denaturation step for 5 min at 95° C. was followed by 40 cycles of PCR (95° C. for 1 min, 55° C. for 1 min, and 72° C. for 2 min). Random PCR products were then separated on a 1.5% agarose gel, and DNA smears ranging in size from approximately 400 to 1,500 by were excised and extracted using the QIAQUICK gel extraction kit (Qiagen). Equal volumes of the eluted, purified PCR products generated from nuclease resistant DNA and the randomly amplified PCR product generated from nuclease resistant RNA were mixed together, ligated into the pGEMT-EASY vector (Promega) and introduced into chemically competent Escherichia coli TOP-10 cells (Topo One Shot; Invitrogen).

[0216] For HBoV and HBoV2 PCR 140 μl of clarified stool supernatant was extracted into 60 μl H₂O using the Qiagen MinElute kit. 12 μl of nucleic acid was used in the PCR. Feces from the UK were tested in pools of ten and then resolved to single sample. HBoV was tested using the nested PCR conditions previously described (Manning et al., J Infect D is 194:1283-90, 2006). HBoV2 was tested using first round PCR primers: HBoV2-sf1 AACAGATGGGCAAGCAGAAC (SEQ ID NO:62) and HBoV2-sr1 AGGACAAAGGTCTCCAAGAGG (SEQ ID NO:63). Second round PCR primers were: HBoV2-sf2 TGCTTCAACAGGCAAAACAA (SEQ ID NO:64) and HBoV2-sr2 TCCAAGAGGAAATGAGTTTGG (SEQ ID NO:65) amplifying a 454 nucleotide region within the NS open reading frame (ORF). PCR were in 50 μl volume at a final MgCl₂ concentration of 1.5 mM, using GOTAQ (Promega), and 0.4 μM of each primer. First round cycling was at 95 C for 45 sec., 55 C for 1 min., 72 C for 1 min. for 5 cycles followed by 95 C for 30 sec., 53 C for 30 sec., 72 C for 45 sec. for 35 cycles. Similar conditions were used for the second round except that the first and second annealing temperatures were 53 C and 50 C. The PCR were analyzed by agarose gel electrophoresis and the positive PCR directly sequenced. The sensitivity of the HBoV2 PCR was measured using dilutions of a NS plasmid subclone. 10, 5 and 1 plasmid copies were detected in 10/10, 8/10 and 2/10 PCR respectively.

[0217] The resulting random PCR products derived from nuclease resistant, viral-sized particles associated RNA and DNA were subcloned and plasmids inserts sequenced. The resulting sequences were analyzed by tBLASTx against Genbank database. The viral 5' and 3' extremities of HBoV2 were amplified using a modification of RACE (rapid amplification of cDNA ends) (Jones et al., J Virol 79:8230-6, 2005). Conditions for HBoV and HBoV2 PCR are described above.

[0218] Sequence distances for different genomic regions were measured using built in functions in the Simmonics2005 sequence editor v1.6. Trees were constructed from pairwise nucleotide and amino acid sequence distances by neighbour-joining in the MEGA2 package. The robustness of groupings was calculated by bootstrap re-sampling of 1000 replicates of the data.

[0219] Viral-sized particles were first purified from two consecutive stool samples from a Pakistani child with AFP. Nuclease resistant (i.e., capsid protected) viral nucleic acids were then extracted and randomly amplified (see above). The resulting random amplification products were subcloned and 97 plasmid inserts were sequenced and analyzed by tBLASTx.

[0220] Exact sequence matches were found in Genbank to human sequences as well as to Micrococcus luteus, Pseudomonas fluorescens and uncultured bacterium sequences. Highly significant but imperfect matches (E score)<10^-10 we also found to Chlamydophila pneumoniae, Rhodoferax ferrireducens and numerous bacteriophages. A single perfect sequence match was found to human poliovirus 1 vaccine strain Sabin 1. The detection of a polio Sabin 1 viral sequence likely reflected ongoing replication of orally administered polio vaccine in this child of 36 months who had previously received a total of 14 oral polio vaccinations.

[0221] 11/47 plasmid sequences from the first stool sample (plus 19/48 from the second time point) gave highly significant tBLASTx E scores to the HBoV genome reference sequence (NC_--007455). PCR was then used to link the different HBoV-like fragments while 5' and 3' RACE were used to amplify the viral extremities. 5196 bases of a novel bocavirus genome were assembled. Based on the ST2 prototype genome sequence of HBoV it is estimated that at least 7 bases may be missing from the 5' end, while the genome described here extends for a further 25 bases at the 3' end. Because the closest genetic relative of this new virus was HBoV, it was termed HBoV2.

[0222] The arrangement of ORF's in the prototypic HBoV2 genome was similar to that of HBoV, with three large, coding sequences (FIG. 13). The 5'NS1 ORF is required for viral DNA replication and the regulation of viral gene expression. Its protein sequence identity with HBoV was 78% and was co-linear through the gene. The second ORF, NP1, was 4 amino acid shorter and 67% identical. NP1 is a protein of unknown function restricted to bocaviruses (Schwartz et al., Virology 302:219-23, 2002; and Chen et al., J Virol 60:1085-97, 1986). The third large ORF encoded a protein with 80% identity to the VP1/VP2 of HBoV. Relative to the HBoV coding sequence, the VP1 of HBoV2 was preceded by a 25 amino acids methionine initiated ORF stretch and a four amino acid deletion downstream resulting in a slightly larger VP1. Through comparison with HBoV, the VP2 protein of HBoV2 was predicted to start at the 154th amino acid of the third large ORF.

[0223] To investigate whether PCR assays for HBoV would be able to amplify HBoV2, the PCR primers from the liteiature used for HBoV (Allander et al., Proc Natl Acad Sci USA 102:12891-12896, 2005; Manning et al., J Infect Dis 194:1283-90, 2006; Sloots et al., J Clin Virol 35:99-102, 2006; Endo et al., J Clin Microbiol 45:3218-23, 2007; Bastien et al., Emerg Infect Dis 12:848-50, 2006; Smuts et al., Emerg Infect Dis 12:1457-8, 2006; Kesebir et al., J Infect Dis 194:1276-82, 2006; Bastien et al., J Clin Microbiol 45:610-3, 2007; Chung et al., Emerg Infect Dis 12:1254-6, 2006; Foulongne et al., Emerg Infect Dis 12:862-3, 2006; Qu et al., Emerg Infect Dis 13:165-8, 2007; and Albuquerque et al., Emerg Infect Dis 13:1756-8, 2007) were aligned with the homologous regions of HBoV2. Most PCR primers contained a substantial number of mismatches with HBoV2 that would preclude or greatly reduce the efficiency of amplification and amplicon detection.

[0224] To determine the relationship of HBoV2 to other members of the Bocavirus genus, phylogenetic analyses were performed for the three large ORFs (VP1/2, NP-1 and NS) using both nucleotide and deduced protein sequences (FIG. 14). The prototype HBoV2 variant (PK5510), while more closely related to HBoV than the animal bocaviruses, CnMV and BPV-1, consistently adopted an outlier position to the Glade containing all the published HBoV sequences in all three genomic regions. Reflecting this, pairwise nucleotide distances between HBoV2 and HBoV were substantially greater (22-26%) than within the HBoV Glade (0.4-0.9%) (Table 1), but less than between HBoV or HBoV2 sequences and animal bocaviruses (46-56%). Similarly, while almost no amino acid sequence variability was observed among HBoV sequences in any genomic region (0.2-0.5%), sequences of all three major genes differed substantially from HBoV2 (20-33%). An unusual partial NS1 HBoV sequence has been reported in Brazil and adopted an intermediate position between HBoV and HBoV2 on phylogenetic analysis (FIG. 14 Partial NS1 EF560212). Interestingly this Brazilian HBoV sequence was derived from the feces of a child with gastrointestinal symptoms (Albuquerque et al., Emerg Infect Dis 13:1756-8, 2007).

TABLE-US-00001 TABLE 1 Nucleotide and Protein Sequence Comparisons Among and Between Bocaviruses. Comparison Region Comps. Nucleotide Amino acid dN/dS A) Main ORFs Within HBoV2: VP1/2 3 3.1% 1.1% 0.094 '' NP1 3 5.9% 8.7% 0.348 '' NS1 3 5.8% 4.3% 0.103 Within HBoV: VP1/2 55 0.9% 0.5% 0.075 '' NP1 55 0.4% 0.3% 0.103 '' NS1 55 0.4% 0.2% 0.068 HBoV-HBoV2: VP1/2 33 22.2% 20.2% <0.13* '' NP1 33 22.9% 32.9% <0.38* '' NS1 33 26.2% 27.8% <0.19* HBoV2-B/CnPV: VP1/2 12 46.2-46.3% .sup. 51.9-53.2% .sup. n.c. '' NP-1 12 47.0-52.3% .sup. 59.7-61.3% .sup. n.c. '' NS1 12 51.3-55.8% .sup. 60.6-65.1% .sup. n.c. B) Partial NSI region (positions 1457-1748) Within HBoV2 genotypes: 7 0.9% 0.3% 0.022 Within HBoV (main group): 55 0.7% 0.0% 0.000 Between HBoV2 genotypes: 21 4.5% 0.6% 0.007 HBoV - EF560212: 11 9.0% 1.1% 0.003 HBoV-HBoV2: 88 18.0% 8.3% 0.071

[0225] To determine the prevalence of human bocaviruses nested PCR primers specifically targeting HBoV and HBoV2 NS region were used. DNA from the stool samples of 57 Pakistani children with AFP and 41 healthy Pakistani children, plus 699 stool samples submitted for enteric bacteriology screening in Edinburgh were analyzed. A total of three AFP stool samples (including the original patient 5510) and two stools from healthy Pakistani children were positive for HBoV2 DNA sequences (ages were 12, 16, 36, 36, and 96 months). HBoV was not found among the 57 AFP stools. The stool samples from the UK were tested in pools of 10 samples in which two HBoV (ages 1-2 years and 3-5 years) and three HBoV2 (ages 0-3 months, 6-12 months and >65 years) positive samples were identified.

[0226] To investigate the genetic diversity of HBoV2, the partial NS region amplicons from the Pakistan and UK samples were sequenced and compared to prototypic HBoV2 and other available human and animal bocavirus sequences (FIG. 14, Partial NS1). Although this initial survey was small, HBoV2 variants fell into 3 groups or genotypes (gts) showing 4.5% sequence divergence from each other in this region of the bocavirus genome (Table 1), a level of diversity much greater than observed between known variants of HBoV (0.7% in this region). HBoV diversity was more comparable to mean diversity seen within any of the three HBoV2 genotype (0.9%).

[0227] To further investigate HBoV2 diversity, near complete genome sequences were obtained of representatives of the other two NS1 based genotypes (gt1: HBoV2 prototype PK5510, gt2: PK2255, and gt3: UK648). Neither HBoV2 variant contained the methionine initiated 25 amino acid stretch seen upstream of VP 1 in PK5510 prototype but had otherwise identical length NS1 and NP1 genes (except from NP1 of PK5510 that was one amino acid shorter). As observed for HBoV, sequence divergence within HBoV2 was highly conservative, with very low dN/dS ratios in all three genome regions (Table 1). Despite the approximately equal degrees of sequence divergence between the HBoV2 genotypes in the partial region of NS1 initially sequenced, phylogenetic relationships varied in different genome regions (FIG. 14). The gt1-PK5510 and gt2-PK2255 variants clustered closely in VP1, with gt3-UK648 as a outlier, while gt3-UK648 and gt2-PK2255 clustered in the NP1 and NS1 genes with gt1-PK5510 as the outlier (FIG. 14). Using sliding window analysis of pair-wise nucleotide distances over the HBoV2 genome, gt2-PK2255 and gt3-UK648 showed a lower level of divergence relative to the other two pair-wise comparisons in the NS1 and NP1 regions (FIG. 15) while gt1-PK5510 and gt2-PK2255 were nearly identical in the VP1/VP2 region (FIG. 15). Discordant phylogenies and inconsistent sequence divergence values between HBoV2 variants is consistent with the occurrence of complex recombination events in the evolution of these viruses. Putative breakpoints were located near the middle and 3' end of NS1 and potentially more recently (i.e., recombinants showing a lower level of divergence) near the beginning of the VP1/VP2.

[0228] Based on the VIIIth report of the International Committee on Taxonomy of Viruses (ICTV), different bocavirus species should show NS gene nucleotide sequence similarities below 95%. HBoV2, showing 75.6% nucleotide similarity to its closest relative HBoV therefore qualifies as a new human parvovirus and the fourth species in the Bocavirus genus following BPV, CnMV and HBoV. It is believed that this terminology is more appropriate than its alternative possible name as HBoV genotype 2 because genotypes of other parvoviruses, such as those reported for B19 and PARV4 are much less divergent from each other and do not qualify as separate species using ICTV guidelines.

[0229] HBoV2 divergence also likely precluded its detection using HBoV based PCR. Thus the epidemiological and clinical information on human bocavirus collected to date refers exclusively to HBoV, and the available sample archives of clinical specimens will have to be screened again with HBoV2-specific primers to investigate its frequency and potential disease associations.

[0230] Because parvoviruses are particularly hard to inactivate using heat and detergent treatment they are of special concern in blood product transfusions and some countries screen for parvovirus B19 DNA to exclude highly viremic donations from blood derived products. The development of serological assays, as recently achieved for HBoV (Kantola et al., Clin Infect Dis 46:540-6, 2008; and Endo et al., J Clin Microbiol 45:3218-23, 2007), will allow larger epidemiological studies to measure the rate of seroconversion in different age and geographic cohorts and whether detection of IgM and rise in IgG titers are associated with particular symptoms.

[0231] Three genotypes of HBoV2 were characterized whose genetic distances to one another (in partial NS1 sequences) were comparable to those measured between B19 or PARV4 genotypes. The HBoV2 genotype geographic distribution appeared different with all three HBoV2 gt3 being derived from the UK while both genotype 1 and 2 where found in Pakistan. Recombination events among HBoV2 variants were also observed as recently reported for animal parvoviruses (Shackelton et al., J Gen Virol 88:3294-301, 2007). The existence of geographic clustering, further genotypes, and different patterns of recombination will require further screening of samples from a wider geographical base.

[0232] If HBoV2 is found archived in tissue as shown for B19 and PARV4 (Manning et al., J Infect Dis 195:1345-52, 2007; Hokynar et al. Virology 302:224-8, 2002; Norja et al., PNAS 103:7450-3, 2006; and Norja et al., J Virol, 2008) the possibility of past epidemic waves of different HBoV2 genotypes will also be testable using tissues from cohorts of different ages.

[0233] Although the detection of HBoV2 was limited to fecal samples in the current study, its detection in stools from 5 Pakistani children lends further supports to the gastrointestinal tract as a site of replication of human bocaviruses. The detection of HBoV2 in the stool of 3 UK residents (2 being less then 12 months old and 1>65 years old) also indicates that this virus is not restricted to South Asia nor to young children.

Sequence CWU 1

6515196DNABocavirus 1ggagtggcta cgtatggggt gatcataaac acgcccagga agtgacgtat gtcaaccaat 60cagcatcgag catatatcct atataagccg atgcactycc gcatctcgtc agactgcatc 120cggtctccgg cgagtgaaca tctctgggaa gagctccacg cacgtggtga gtgacactat 180ggccttttct gctcctgtaa ttagagcttt ttctcaacct gcttttactt atgttgttaa 240atttccatat gaaaactgga aagaggaaga gcacttacta tggagcttac ttgctcctgg 300gactgaacgt ctcatgattc aactaagaaa ctgcgcacca catcctgaag atgatcctgt 360cagggaagat attttatgct cactagcaga tcaacactat ggtgctattt ttgctaaagc 420ttgctacata gctacaacta cactaatggg gcagaaacag agaacacctt ttccacgctg 480cgacattata tgccaatctg aaattggttc agaacatcta cattgtcaca tacttgttgg 540aggagccggt ctcagcaaaa gaaatgctaa aatttcacgt gctacacttc tgggtcttgt 600gatggctgaa ctaacacaac gctgcaaaca gcttcttgcg cttcgtccat ttgaaccagc 660tgaggctaat atttttcatc tcctcaagcg cattgaacgc gaagcatggt cagggcatac 720tggtaactgg gttcaaattc ttcaatacaa agataagcga ggtgatcttc acgctcaacc 780aattgatcct ttacgctttt taaagcatta catactacca aaaaatcgat tgatttctcc 840ttccagcaaa cctgacgtct gcactactcc agataactgg ttcattctag ctgacaaaac 900atacgctcac actattatta atgggcttcc gctgctagaa cataacagaa aagcctatct 960acaagagtta gaaagtgaag tcatcccggg gccttctacc atggcctttg ggggacgtgg 1020tgcgtgggaa caacttcctg aggtaggaga acaacgctta attacttcta atgcttctac 1080tgcttataaa gctaacaaaa aagaaaaatt aatgctaaat ttacttgata aatgtgatga 1140acttaattta cttgtttatg aagacttagt tagtgcttgt cctgaccttt tacttatgct 1200tgaaggtcag ccaggtggtg cacgcctaat tgaacaggtg cttggcatgc atcatattaa 1260ggtgtgtgct aaatacactg cattaacatt tttatttcat ttacatccgg atcaattatt 1320aacttctaac aataaagctt taaaactact gttgattcaa gggtacaacc cacttcaagt 1380agggcatgcc atctgttgtg tacttaacaa acagatgggc aagcagaaca ctatctgctt 1440ttatggtcct gcttcaacag gcaaaacaaa ctttgcaaaa gcaatagttc aaggtgttcg 1500cctttatggc tgtgttaatc atttaaacaa agggtttgtc tttaacgatt gcagacaacg 1560ccttataatt tggtgggaag agtgtttaat gcaccaagat tgggtggaac ctgctaaatg 1620cattttaggc ggaactgaat gtagaattga tgttaaacat aaagacagtg ttcttcttca 1680acaaacacca gtaattattt ccactaacca tgacatctac tctgtagttg gtggcaatac 1740tgtttctcat gttcatgcag caccattaaa agagcgagtc cttcagctaa atttcatgaa 1800acaactgcca caaacatttg gagaaatttc tccagttgaa attgcagaac tattgcaatg 1860gtgctttaat gagtacgaat gtactcttac tggctttaaa caaaaatgga acttagataa 1920agttccaaac tcatttcctc ttggagacct ttgtcctaca cattcacagg actacacgct 1980tcacgaaaac ggattctgca ctgactgtgg cggctatctt cctcatagtg ctgacgattt 2040tgtctacact gacgtggcta gcgaaacaac aagcggagac tgcgacccag gtaggcttta 2100atacatttgc ttaattaaat tatatatttg cactttgctt atgtattaac tcctacaggt 2160aacctggggg atacggacgg agaggactcc aagtcagagg catcggaagt ggactttcgt 2220ccatccaaga agaggcgtgt gatttcagca actccaccaa gcagtccagt aagtggtcca 2280agcctttcta cctttttaga tacttggcag tcacaaccta gggacgaaga tgagctccga 2340atctatgaag aacaagcatc gcaactccaa aagaacacca agtccactcc agaaagagag 2400gaggcgcaac tgggagagcc acaagagccg cagccggagc ccgatccgac ggcatgggga 2460gaaaaacttg gagtatgctc atcacagcaa ccaggagaac cgccagtcgt cttatactgc 2520tttgaagacc tcagaccaag cgacgaagac gaaggagaaa acatcggggg ggaatagaac 2580caatccttat actgtattca gtcaacacag ggctaatcat tcagatgctc ctggatggtg 2640tgggttttac tggcattcta ctagacttgc tagagatggg actaattgta tctttaatga 2700aatgaaacaa gaatttcaag aattacaaat aaatggaaaa attacttggg acaatgctag 2760agaactattg tttagtcaga aaaaaaagct agatcaaaaa tacagaaaca tgctgtatca 2820tttcagacac agtcctgatt gtcctagatg tgattattgg gataatgtat accgtagaca 2880cttagctcat gtctcttcac aggaatcaga ggaggttaca gacgaagaaa tgctttctgc 2940tgttgaaagc atggatacaa atgcctccaa ttaaacgcca gcctggaggg tgggtgcttc 3000ctggttataa ataccttggt ccattcaatc ctcttgaaaa cggtgaacca gttaataaag 3060ctgatcgtgc tgctcaagct catgataaat catattctga aataataaaa agtggaaaaa 3120atccttactt gtatttcaat aaagctgatg aaaaattcat tgacgatttg aaaaacgact 3180ggtctcttgg tggcattatt ggctcaagtt tctttaaact taagcgcgcc gtggctcctg 3240ctctaggaaa taaagagcga gctcaaaaaa gacattttta ctttgcaaac tcaaataaag 3300gtgctaaaaa accaaaaaat aacgagccta aaccaggcac atcaaaaatg tctgaaaatg 3360aaatccaaga ccaacaacca tctggctcca tggaggagcg aggaggcgga ggaggtgcgg 3420tcggtagtgt gggagggggg aaaggttcta gtgtgggtat atccacaggc ggctgggttg 3480gaggcagcta ctttactgac tcatatgtca taacaaagaa cactagacag ttcttagtta 3540aaatacaaaa tgaccacaaa tacagaacag agaatataat tccaagcaac gcaggaggaa 3600aatcccagcg atgcgtaagc acaccttggt catactttaa cttcaatcaa tacagcagtc 3660acttctcacc acaagactgg cagcatttaa caaatgaata taaacgcttt aagcctagaa 3720aaatgcatgt aaaaatttac aacttacaaa taaaacaaat actctcaaat ggtgctgaca 3780ctacatacaa caacgaccta acagctggtg ttcacatctt ttgtgatggt gaacacgcat 3840atccaaatgc aacacatcca tgggatgaag atgtcatgcc agaacttcca tatgaaacat 3900ggtatttgtt tcaatatgga tacattccag ttattcatga actggctgaa atggaagacg 3960caaatgctgt agaaaaagct atagcactac aaataccctt tttcatgctt gaaaacagcg 4020accatgaagt gttaagaaca ggagaaagca cagaattcac ttttgacttt gactgtgaat 4080ggataaacaa cgaaagagca tacattcctc ctggattaat gtttaatcca aaagttccta 4140caagaagagc tcaatacatc agacagcacg gaaacacagc atccagcaac accagaattc 4200aaccatatgc aaaacctaca agctggatga caggaccagg tctactcagc gcacaaagag 4260taggaccagc tggctcagac actgcatcat ggatggttgt tgtcaatcca gacggagctg 4320cagttaactc aggaatggca ggagttggtt caggatttga tcctccttca ggatctctaa 4380gaccaactga cttagaatac aaaatacaat ggtaccaaac tcctgcaggt accaacagtg 4440atggaaacat catttcaaat ccaccactat ccatgctcag agatcaagct ctctacagag 4500gaaatcaaac aacctacaac ctatgctcag atgtgtggat gttcccaaat caaatttggg 4560acagatatcc aataaccaga gaaaatccaa tctggtgtaa aaaaccaaga tcagacaaaa 4620acacaataat tgatcctttc gatggaacac ttgcaatgga tcatccgcca ggaacaatct 4680tcataaaaat ggcaaaaatt ccagttcctt caaacaacaa cgcagactca tacctaaaca 4740tctactgcac tggacaagtc agctgcgaaa ttgtctggga agttgaaaga tacgcaacaa 4800agaactggag accagaaaga agacacaccg cacttggtct tggaattgga ggagaagaaa 4860acgtaaatcc aacttatcat gtagacaaaa atggaaaata cattcaacca acaacttggg 4920acatgtgcta tcctatcaaa acaaacatca ataaagtgtt gtaatctctt aagcctgttc 4980attgcttatg cttataagtt cctctccaat ggacaagagg aaagaaaagg gtgactgtaa 5040tcccgagctc atgagttcga ggctacagtc cgatggcagt ggtgttgccg tctcgaacct 5100agccggtaca cccttgtgca ttgtgggagg agctgttttg cttacgcaat cgcgaaattt 5160tatatattta atgtagtgtt gtactgcgtc aggcat 51962640PRTBocavirus 2Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Glu Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Gly Ala Ile Phe Ala Lys65 70 75 80Ala Cys Tyr Ile Ala Thr Thr Thr Leu Met Gly Gln Lys Gln Arg Thr 85 90 95Pro Phe Pro Arg Cys Asp Ile Ile Cys Gln Ser Glu Ile Gly Ser Glu 100 105 110His Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Arg Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Gln Leu Leu Ala Leu Arg Pro Phe Glu Pro145 150 155 160Ala Glu Ala Asn Ile Phe His Leu Leu Lys Arg Ile Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Lys Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln Pro Ile Asp Pro Leu Arg Phe Leu 195 200 205Lys His Tyr Ile Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Ile Leu Ala Asp Lys225 230 235 240Thr Tyr Ala His Thr Ile Ile Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu Gln Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Ala Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Lys Tyr Thr Ala 355 360 365Leu Thr Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Asn 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Phe 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Thr Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Leu Pro His Ser Ala Asp Asp Phe Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Thr Ser Gly Asp Cys Asp Pro Gly Arg Leu625 630 635 6403214PRTBocavirus 3Met Ser Ser Glu Ser Met Lys Asn Lys His Arg Asn Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Gln Lys Glu Arg Arg Arg Asn Trp Glu Ser His Lys 20 25 30Ser Arg Ser Arg Ser Pro Ile Arg Arg His Gly Glu Lys Asn Leu Glu 35 40 45Tyr Ala His His Ser Asn Gln Glu Asn Arg Gln Ser Ser Tyr Thr Ala 50 55 60Leu Lys Thr Ser Asp Gln Ala Thr Lys Thr Lys Glu Lys Thr Ser Gly65 70 75 80Gly Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala Asn 85 90 95His Ser Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr Arg 100 105 110Leu Ala Arg Asp Gly Thr Asn Cys Ile Phe Asn Glu Met Lys Gln Glu 115 120 125Phe Gln Glu Leu Gln Ile Asn Gly Lys Ile Thr Trp Asp Asn Ala Arg 130 135 140Glu Leu Leu Phe Ser Gln Lys Lys Lys Leu Asp Gln Lys Tyr Arg Asn145 150 155 160Met Leu Tyr His Phe Arg His Ser Pro Asp Cys Pro Arg Cys Asp Tyr 165 170 175Trp Asp Asn Val Tyr Arg Arg His Leu Ala His Val Ser Ser Gln Glu 180 185 190Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser Met 195 200 205Asp Thr Asn Ala Ser Asn 2104667PRTBocavirus 4Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Ile 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Leu Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Pro Lys Asn Asn Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Gly Ser Met 130 135 140Glu Glu Arg Gly Gly Gly Gly Gly Ala Val Gly Ser Val Gly Gly Gly145 150 155 160Lys Gly Ser Ser Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 165 170 175Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 180 185 190Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro 195 200 205Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 210 215 220Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp225 230 235 240Gln His Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His 245 250 255Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 260 265 270Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys 275 280 285Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 290 295 300Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly305 310 315 320Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala 325 330 335Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 340 345 350Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe 355 360 365Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 370 375 380Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile385 390 395 400Arg Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr 405 410 415Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 420 425 430Arg Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val 435 440 445Asn Pro Asp Gly Ala Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser 450 455 460Gly Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr465 470 475 480Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly Asn 485 490 495Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr 500 505 510Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe 515 520 525Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 530 535 540Trp Cys Lys Lys Pro Arg Ser Asp Lys Asn Thr Ile Ile Asp Pro Phe545 550 555 560Asp Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 565 570 575Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 580 585 590Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val 595 600 605Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 610 615 620Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr His625 630 635 640Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 645 650 655Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 660 66554972DNABocavirus 5gtggtgagtc acattatggc tttcaatcca cctgttatta gagcattctc ttcacctgct 60tttacttatg tcttcaaatt tccatatcca tcatggaaag aaaaagaatg gcttcttcat 120gcacttctgg ctcatggtac cgagcaagcc atgatccagc tgagaaactg tgttcctcat 180ccggatgaag atataatccg tgatgactta ctactttctc tagaagatcg ccattttggg 240gcaattctct gcaaggctgt ctatatggct actactactt ttatgtcaca gaaacaaaga 300aatatgtttc ctcgctgtga cataatagtc caatctgagc ttggggagac aaacctacac 360tgccatatta tagttggggg agaaggctta agcaagagaa atgcaaaaac atcatgtcct 420caactatatg gactgatact aggggaatta atccaacgct gcaaaactct tctggctacg 480cgtccttttg aaccggaaga ggcagaaatt tatcatgctt

taaaacgagc tgagcgagaa 540gcttggggtg gagttactag cggcaaccta caaattctcc aatacagaga tcgcagagga 600gaccttcacg cacaacaagt ggatgctctt cgcttcttca aaaactacct attgcctaaa 660aatagatgca ttacatctta cagcagacct gatgtctgta cttctccaga aaactggttt 720gttttagctg aaaaaactta ctgtcacact cttgttaacg ggctgccgct tccagaacat 780tacagaaaac actaccacgc aaccctagat aacgaagttc taccagggcc tcagacaatg 840gcctttgggg gacgtggtcc gtgggaacat cttcctgagg taggagatca acgtttagct 900gcttcttctg ttagtacaac atataaacca aacaaaaaag aaaaacttat gcttaactta 960ctagataaat gcagcgaatt aaatctttta gtttatgaag acttagtagc taactgtcct 1020gaacttttgc ttatgcttga aggtcaacca ggtggggcac gcttaataga acaagtccta 1080ggcatgcacc atattaatgt ttgctctaac tttactgctc ttagttatct ctttcacctt 1140taccctggca caaccttatc ttcagataac aaggctttgc agctgttgtt gatacaaggt 1200tacaacccat taatggttgg tcacgccttg tgctgtgtac tcaacaagca atttggcaaa 1260caaaacactg tttgctttta tggaccagct tctactggta aaacaaacat ggcaaaggcc 1320atagtccaag gcattagact atatggctgt gttaatcatt taaacaaagg gtttgtcttt 1380aatgattgca gacaacgcct agttgtttgg tgggaggagt gcttaatgca ccaggattgg 1440gtggaaccag caaagtgtat cttgggtgga actgagtgta gaattgacgt caaacacaga 1500gatagtgtat tattgacaca aactccagta attatttcca ctaaccacga tatctacgcg 1560gttgttggtg gtaattctgt ttctcatgtt catgcggctc cattaaaaga aagagtgatt 1620cagctaaatt ttatgaaaca acttcctcaa acatttggag agatcactcc agaagaaatt 1680gcagctctac tgcaatggtg tttcaatgag tacgaatgta ctctgacagg ctttaaaaca 1740aaatggagcc tagataaaat tccaaactca tttcctcttg gggtcctttg tcctactcat 1800tcacaggact tcatactcca cgaaaacgga tactgcactg attgtggtgg ttaccttgct 1860catagcgctg acgattctgt gtacactgat cgtgcaagcg acactagcaa agaagccatc 1920gacgcaggta agtttacgtt ctccaggcac tttatatata tcctatacac aacactaaaa 1980catatgttta attacaggtg acttggggga tacggacgga gaggactccg agtcagaagc 2040atcggaagtg ggtgttcgtc catccaagaa gcgacgcata actattcctg caactccacc 2100aaattctcct ggcagctctg tgagtacttc tgccttcttt gataattggt gcgcacaacc 2160gcgagacgaa gatgagctca gggaatacga aagacaagca tcgcgcctac aaaagaaaag 2220ggagtccagg gagagacgag aagaaacgcc catggcaacc tcgtcacagg agtcggagtc 2280ggagcccaat ccgacgcagt ggggagacaa gctcggggtc ataccgtcag gaacaccaga 2340tcagccacct atcgtcttgc actgcttcga agatctcaga cccagtgacg aagacgaagg 2400agaatacatc gggaaagaga gactctagaa ctaatccata cactgtattc agccagcata 2460aagcctcaca tcctgatgct ccaggatggt gtgggttcta ttggcattct actagaattg 2520ctagaaatgg tactaatgca atctttaatg aaatgaaaca gcagttccaa caactgcagc 2580tagacaacaa aattggctgg gataatgcta gagaattatt gtttagtcag aaaaaatcac 2640tagatcaaca atacagaaat atgttctggc actttagaaa tgcttctgat tgtgaacgtt 2700gtaattactg ggacaatgta taccgtatgc acttagctca tgtttcctct cagacagaat 2760cagaagaaat aactgacgag gaaatgcttt ctgctgctga aagtatggaa acagatgcct 2820ccaattaaaa ggcaacctgg agggtgggtg cttcctggtt acaaatacct tggtccattt 2880aatcctcttg ataacggtga accagttaat aaagctgatc gtgctgctca atctcatgat 2940aaatcatatt ctgaattaat aaaaagtgga aaaaatcctt acttatattt caataaagct 3000gatgaaaaat tcattgacga tttgaaaaac gactggtctc ttggtggcat tattggctca 3060agtttcttta aacttaagcg cgccgtggct cctgctctag gaaataaaga gcgagctcaa 3120aaaagacact tttattttcc aaactcaaat aaaggtgcta aaaaatcaaa aaacaacgaa 3180cctaaaccaa acacctcaaa aatgtctgaa aatgaaattc aagatcaaca gccatcagaa 3240cctaatgatg gccaacgagg agggggagga ggtgcgaccg gcagtgtggg aggggggaaa 3300ggttctggtg tgggtatatc cacaggtgga tgggtaggag gcagctactt tactgactcc 3360tatgtcataa caaaaaacac cagacaattt ctggttaaaa tccaaaacaa ccatcaatat 3420aaaactgaaa atataattcc ttccaatgga ggaggaaaat cacaaagatg tgtcagcaca 3480ccatggtcat actttaactt taatcaatac agcagtcatt tctcaccaca ggactggcag 3540cgcctaacaa atgaatacaa aagattcaga cctaaaggta tgcatgttaa aatctacaat 3600ttacaaataa aacagatttt atcaaatggt gctgatgtta catacaacaa cgacctaaca 3660gcaggagtac acatcttttg tgatggcgaa catgcatatc caaacgctac acatccatgg 3720gacgaagatg taatgccaga acttccttac caaacatggt atctgtttca atatggatac 3780atacctacca ttcatgaact tgcagaaatg gaagactcca atgcagtaga aaaagcaatt 3840gctttacaga taccattctt catgcttgaa aacagcgacc atgaagttct aagaactgga 3900gaaagtgcag aatttaactt caactttgac tgtgaatgga ttaacaatga aagagcattc 3960attcctccag gactgatgtt taatccattg gtaccaacaa gaagagctca atacatacga 4020agaaatggaa acactcaagc aagtacatca cgaattcaac cctatgctaa acctacaagc 4080tggatgactg ggccaggttt actcagtgca caacgagtag gtccagctgc ttctgacaca 4140gctgcatgga tggttggtgt agatccagaa ggcgcaaaca tcaactcagg aagagcagga 4200gttagcagtg gatttgatcc tccagctgga tcactcagac ctacagatct agaatacaaa 4260gtacaatggt accaaactcc agctggaaca aacaacgatg gaaacatcat ttcaaatcca 4320cctttatcaa tgcttagaga tcaaactctc tacagaggaa accaaacaac ctacaactta 4380tgctcagatg tatggatgtt tccaaatcaa atttgggaca gatacccagt aacaagagaa 4440aatcctattt ggtgcaaaca accaagatca gacaaacaca caacaattga tccttttgac 4500ggatcaatag ccatggatca tccaccaggc acaattttca tcaaaatggc aaaaattcca 4560gttccttcaa acaacaacgc agactcatac ttaaacatct actgcactgg acaagtcagc 4620tgcgaaattg tctgggaagt cgaaagatat gcaacaaaga actggagacc agaaagaaga 4680cacacagcac tcggccttgg aattggaggg gcagatgaaa tcaacccaac ataccatgtt 4740gacaaaaacg gagcatacat tcaacctaca acatgggaca tgtgctttcc agttaaaaca 4800aacattaata aagtgttgta atctcttaag cctctttatt gcttacgctt gtaagttcct 4860ctccaatgga caagtggaaa gaaaagggtg actgtaatcc cgagctcatg agttcgaggc 4920tacagtccga tggcagtggt gttgccgtct cgaacctagc cgttacaccc tt 49726661PRTBocavirus 6Met Ala Phe Asn Pro Pro Val Ile Arg Ala Phe Ser Ser Pro Ala Phe1 5 10 15Thr Tyr Val Phe Lys Phe Pro Tyr Pro Ser Trp Lys Glu Lys Glu Trp 20 25 30Leu Leu His Ala Leu Leu Ala His Gly Thr Glu Gln Ala Met Ile Gln 35 40 45Leu Arg Asn Cys Val Pro His Pro Asp Glu Asp Ile Ile Arg Asp Asp 50 55 60Leu Leu Leu Ser Leu Glu Asp Arg His Phe Gly Ala Ile Leu Cys Lys65 70 75 80Ala Val Tyr Met Ala Thr Thr Thr Phe Met Ser Gln Lys Gln Arg Asn 85 90 95Met Phe Pro Arg Cys Asp Ile Ile Val Gln Ser Glu Leu Gly Glu Thr 100 105 110Asn Leu His Cys His Ile Ile Val Gly Gly Glu Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Thr Ser Cys Pro Gln Leu Tyr Gly Leu Ile Leu Gly Glu 130 135 140Leu Ile Gln Arg Cys Lys Thr Leu Leu Ala Thr Arg Pro Phe Glu Pro145 150 155 160Glu Glu Ala Glu Ile Tyr His Ala Leu Lys Arg Ala Glu Arg Glu Ala 165 170 175Trp Gly Gly Val Thr Ser Gly Asn Leu Gln Ile Leu Gln Tyr Arg Asp 180 185 190Arg Arg Gly Asp Leu His Ala Gln Gln Val Asp Ala Leu Arg Phe Phe 195 200 205Lys Asn Tyr Leu Leu Pro Lys Asn Arg Cys Ile Thr Ser Tyr Ser Arg 210 215 220Pro Asp Val Cys Thr Ser Pro Glu Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Cys His Thr Leu Val Asn Gly Leu Pro Leu Pro Glu His Tyr 245 250 255Arg Lys His Tyr His Ala Thr Leu Asp Asn Glu Val Leu Pro Gly Pro 260 265 270Gln Thr Met Ala Phe Gly Gly Arg Gly Pro Trp Glu His Leu Pro Glu 275 280 285Val Gly Asp Gln Arg Leu Ala Ala Ser Ser Val Ser Thr Thr Tyr Lys 290 295 300Pro Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Ser305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ala Asn Cys Pro Glu 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Asn Val Cys Ser Asn Phe Thr Ala 355 360 365Leu Ser Tyr Leu Phe His Leu Tyr Pro Gly Thr Thr Leu Ser Ser Asp 370 375 380Asn Lys Ala Leu Gln Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Met385 390 395 400Val Gly His Ala Leu Cys Cys Val Leu Asn Lys Gln Phe Gly Lys Gln 405 410 415Asn Thr Val Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Met 420 425 430Ala Lys Ala Ile Val Gln Gly Ile Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Val Val 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Arg Asp 485 490 495Ser Val Leu Leu Thr Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ala Val Val Gly Gly Asn Ser Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Ile Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Thr Pro Glu Glu Ile Ala Ala Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Thr Lys 565 570 575Trp Ser Leu Asp Lys Ile Pro Asn Ser Phe Pro Leu Gly Val Leu Cys 580 585 590Pro Thr His Ser Gln Asp Phe Ile Leu His Glu Asn Gly Tyr Cys Thr 595 600 605Asp Cys Gly Gly Tyr Leu Ala His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Arg Ala Ser Asp Thr Ser Lys Glu Ala Ile Asp Ala Gly Lys Phe625 630 635 640Thr Phe Ser Arg His Phe Ile Tyr Ile Leu Tyr Thr Thr Leu Lys His 645 650 655Met Phe Asn Tyr Arg 6607218PRTBocavirus 7Met Ser Ser Gly Asn Thr Lys Asp Lys His Arg Ala Tyr Lys Arg Lys1 5 10 15Gly Ser Pro Gly Arg Asp Glu Lys Lys Arg Pro Trp Gln Pro Arg His 20 25 30Arg Ser Arg Ser Arg Ser Pro Ile Arg Arg Ser Gly Glu Thr Ser Ser 35 40 45Gly Ser Tyr Arg Gln Glu His Gln Ile Ser His Leu Ser Ser Cys Thr 50 55 60Ala Ser Lys Ile Ser Asp Pro Val Thr Lys Thr Lys Glu Asn Thr Ser65 70 75 80Gly Lys Arg Asp Ser Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His 85 90 95Lys Ala Ser His Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His 100 105 110Ser Thr Arg Ile Ala Arg Asn Gly Thr Asn Ala Ile Phe Asn Glu Met 115 120 125Lys Gln Gln Phe Gln Gln Leu Gln Leu Asp Asn Lys Ile Gly Trp Asp 130 135 140Asn Ala Arg Glu Leu Leu Phe Ser Gln Lys Lys Ser Leu Asp Gln Gln145 150 155 160Tyr Arg Asn Met Phe Trp His Phe Arg Asn Ala Ser Asp Cys Glu Arg 165 170 175Cys Asn Tyr Trp Asp Asn Val Tyr Arg Met His Leu Ala His Val Ser 180 185 190Ser Gln Thr Glu Ser Glu Glu Ile Thr Asp Glu Glu Met Leu Ser Ala 195 200 205Ala Glu Ser Met Glu Thr Asp Ala Ser Asn 210 2158668PRTBocavirus 8Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Asp Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ser His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Leu Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Pro Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Ser Lys Asn Asn Glu Pro Lys Pro Asn Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Glu Pro Asn 130 135 140Asp Gly Gln Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly145 150 155 160Gly Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly 165 170 175Ser Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe 180 185 190Leu Val Lys Ile Gln Asn Asn His Gln Tyr Lys Thr Glu Asn Ile Ile 195 200 205Pro Ser Asn Gly Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp 210 215 220Ser Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp225 230 235 240Trp Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Gly Met 245 250 255His Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly 260 265 270Ala Asp Val Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe 275 280 285Cys Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu 290 295 300Asp Val Met Pro Glu Leu Pro Tyr Gln Thr Trp Tyr Leu Phe Gln Tyr305 310 315 320Gly Tyr Ile Pro Thr Ile His Glu Leu Ala Glu Met Glu Asp Ser Asn 325 330 335Ala Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu 340 345 350Asn Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Ala Glu Phe Asn 355 360 365Phe Asn Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Phe Ile Pro 370 375 380Pro Gly Leu Met Phe Asn Pro Leu Val Pro Thr Arg Arg Ala Gln Tyr385 390 395 400Ile Arg Arg Asn Gly Asn Thr Gln Ala Ser Thr Ser Arg Ile Gln Pro 405 410 415Tyr Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala 420 425 430Gln Arg Val Gly Pro Ala Ala Ser Asp Thr Ala Ala Trp Met Val Gly 435 440 445Val Asp Pro Glu Gly Ala Asn Ile Asn Ser Gly Arg Ala Gly Val Ser 450 455 460Ser Gly Phe Asp Pro Pro Ala Gly Ser Leu Arg Pro Thr Asp Leu Glu465 470 475 480Tyr Lys Val Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Asn Asp Gly 485 490 495Asn Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Thr Leu 500 505 510Tyr Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met 515 520 525Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Val Thr Arg Glu Asn Pro 530 535 540Ile Trp Cys Lys Gln Pro Arg Ser Asp Lys His Thr Thr Ile Asp Pro545 550 555 560Phe Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile 565 570 575Lys Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr 580 585 590Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu 595 600 605Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr 610 615 620Ala Leu Gly Leu Gly Ile Gly Gly Ala Asp Glu Ile Asn Pro Thr Tyr625 630 635 640His Val Asp Lys Asn Gly Ala Tyr Ile Gln Pro Thr Thr Trp Asp Met 645 650 655Cys Phe Pro Val Lys Thr Asn Ile Asn Lys Val Leu 660 66594966DNABocavirus 9tggtgagtga cactatggcc ttttctgctc ctgtaattag agctttttct caacctgctt 60tcacttatgt tgttaaattt ccatatgata actggaaaga ggaagaacac ttactatgga 120gcttacttgc tcctgggact gaacgtctca tgatccaact aagaaactgc gcaccacatc 180ctgaagatga tcctgtcagg gaagatattt tatgctcact agcagaccaa cactatgctg 240ctattttcac caaggcttgc tacatggctg taacttcact catggggcag aaacagagaa 300cacactttcc acgatgcgac ataatatgcc aggctgagat cggctcagaa tatctacact 360gtcacatact tgttggagga gcaggtctga gcaagagaaa tgctaaaatt tcatgtgcta 420cgctcttagg ccttgtgatg gctgaattaa cacaacgctg caaactactt cttacacagc 480gtccatttga accagatgaa gctagaatat tccatctact cagacgcgtt gaacgcgaag 540catggtcagg gcacactggt aactgggttc aaattcttca atacagagac aagcgaggtg 600accttcatgc tcaacacatt gatcctttac gctttttcaa acactacctg ctgccaaaaa 660atcgattgat ctctccttcc agcaagcctg acgtctgcac tactccagat aactggtttg 720tcctagctga taaaacatac gctcacacta ttgttaatgg gcttccgctg ctagaacata 780acagaaaggc ctatctacaa gagttagaga gtgaagtcat cccggggcct tctaccatgg 840cctttggggg acgtggtgcg tgggaacatc tgcctgaggt aggagaacaa cgcctaatta 900cttctaatac ttctactgct tataaagcta acaaaaaaga aaaattaatg ctaaacttac 960ttgataaatg tgatgaactt aacttacttg tgtatgaaga cttagttagt gcttgtcctg 1020accttttact tatgcttgaa ggtcaaccag gtggtgcacg cctaattgaa caggtgcttg 1080gcatgcatca tattaaagta tgtgctaatt acacagcatt atctttctta tttcatttac

1140atcctaatca attattaact tctagcaata aagcactaaa actattgttg attcaagggt 1200acaacccatt acaggtaggg cacgccatct gctgtgtact taacaaacag atgggcaagc 1260agaacactat ctgcttttat ggtcctgctt caacaggtaa aacaaatatt gcaaaggcca 1320tagtccaagg cgttcgcctt tatggctgtg ttaatcatct aaacaaaggg tttgtcttta 1380acgattgcag acaacgcctt ataatttggt gggaggaatg tttaatgcat caagattggg 1440ttgaacctgc taaatgcatt ttaggtggaa ctgaatgtag aattgatgtt aaacacaaag 1500acagtgttct tcttcaacaa acaccagtaa ttatttccac taaccatgac atctactctg 1560tagttggtgg caatactgta tctcatgttc atgcagcgcc cttaaaagag cgaattcttc 1620aactaaattt tatgaaacaa ctgccacaaa catttggaga gatttctcca gttgaaattg 1680cagagttgct gcaatggtgc tttaatgagt acgaatgtac tcttactggc tttaaacaaa 1740aatggaactt agataaagtt ccaaactcat ttcctcttgg ggacctttgt cctacacatt 1800cacaggacta cgtgcttcac gaaaacggat tctgcactga ctgcggcggc tatattcctc 1860atagtgctga cgactctgtg tatactgacg tggctagcga gacatcaatc agcagctgcg 1920acccaggtag gcattaatac attagccttt taatatacta ctttctagat gcttatgtat 1980taactcctac aggtgacttg ggggatacgg acggagagaa ctcccagccg gagacatcga 2040acgtggataa tcgtccatcc aagaagcgac gtgtgattcc agaaactcca ccaaacagtc 2100cagtaagtcg ccaaagcctt tctagctttt tagatacgtg gcagtcacaa cctagagacg 2160aagatgagct ccgaatctat gaagcacagg catcgcgcat caaagagaac accgagtcca 2220ctccggagag agagaagaca ccagtgggag aaccacaaga agagtcgcag gcggagcccg 2280atccgacagc atggggagaa aagcttggag tctactcctc gctgcaacca ggagagccgc 2340caatcgtctt acactgcttc gaagacctca gaccaagcga cgaagacgaa ggagaaaaca 2400tcggggggga atagaaccaa tccttatact gtgttcagtc aacacagggc taatcatcca 2460gatgctcctg gatggtgtgg gttttactgg cattctacta ggcttgctag agatggcact 2520aattgtatct ttaatgaaat gaaacaagaa tttcaagaat tgcaaataaa tgggaaaatt 2580acctgggaca atgttagaga actattgttt agccagaaaa aaaagctaga tcaaaaatac 2640agaaacatgc tgtaccattt cagacataat gctgattgtc ctagatgtga ttattgggat 2700gatgtctacc gtaaacactt agctcatgtc tcttcacagg aatcagagga ggtaacagac 2760gaagaaatgc tttctgctgt tgaaagcatg gaaacaaatg cctccaatta aacgccaacc 2820tggagggtgg gtgcttcctg gttataaata ccttggtcca tttaatcctc ttgagaacgg 2880tgaaccagtt aataaagctg atcgtgctgc tcaagctcat gataaatcat attctgagtt 2940aataaagagt ggaaaaaatc cttacttgta tttcaataaa gctgatgaga aattcattga 3000cgatttgaaa aacgactggt ctcttggtgg cattattggc tcaagtttct ttaaacttaa 3060gcgcgccgtg gctcctgctc taggaaataa agagcgagct caaaaaagac atttttactt 3120tgcaaactca aataaaggtg ctaaaaaacc aaaaaataac gagcctaaac caggcacttc 3180aaaaatgtct gaaaatgaaa tccaagacca acaaccatct ggctcgatgg acgaacagcg 3240aggaggcgga gggggcgccg ttggcagtgt gggagggggg aaaggttctg gtgtgggtat 3300atccacaggt ggctgggtag gaggcagcta tttcactgac tcatatgtca taacaaaaaa 3360caccagacaa tttctggtaa aaatacaaaa tgaccacaaa tacagaactg aaaatattat 3420tccaagcaat gctggaggaa aatcacaaag gtgcgtcagc acaccatggt catatttcaa 3480cttcaatcaa tacagcagtc atttttcacc acaagactgg caacgcctaa caaatgaata 3540taaacgcttt aaacctagaa aaatgcatgt aaaaatttac aatctacaaa taaaacaaat 3600actttcaaat ggtgctgaca ctacatacaa caacgaccta acagctggtg ttcacatatt 3660ttgtgatggt gaacacgcat atccaaatgc aacacatcca tgggatgaag acgtcatgcc 3720agaacttcca tatgaaacat ggtatctgtt tcaatatgga tacattccag ttattcatga 3780acttgctgaa atggaagacg caaatgctgt agaaaaagct atagcactac aaataccatt 3840cttcatgctt gaaaacagtg accatgaagt tctaagaact ggagaaagca cagaattcac 3900ttttgatttt gactgtgagt ggatcaacaa cgaaagagca tacattcctc ctggattaat 3960gtttaatcca aaagttccta cgagaagagc tcaatacatt agacagcacg gaaacacagc 4020atcaagcaac accagaattc aaccatatgc aaaacctaca agctggatga caggaccagg 4080tctactcagt gcacaaagag taggaccagc tggctcagac actgcatcat ggatggttgt 4140tgttaatcca gacggaactg ccgttaactc aggaatggca ggagttggat caggatttga 4200tcctccttca ggatctctaa gaccaactga cttagaatac aaaatacaat ggtaccaaac 4260tcctgaaggt accaacagtg atggaaacat aatttcaaat ccaccactgt ccatgcttag 4320agatcaagct ctctacagag gaaatcaaac aacctataac ctatgctcag atgtatggat 4380gttcccaaat caaatttggg acagatatcc aataaccaga gaaaatccaa tctggtgcaa 4440aaaaccaaga tcagataaaa gcacagtaat tgatcctttc gatggaacac tcgcaatgga 4500tcatcctcct ggaacaatct tcataaaaat ggcaaaaatt ccagttcctt caaacaacaa 4560cgcagactca tacctaaaca tctactgcac aggacaagtc agctgcgaaa ttgtctggga 4620agttgaaaga tacgcaacaa agaactggag accagagaga agacacaccg cacttggtct 4680tggaatcgga ggagaagaaa acataaatcc aacttaccat gtagacaaaa atggaaaata 4740cattcagcca acaacatggg acatgtgcta tcctatcaaa acaaacatca ataaagtgtt 4800gtaatctctt aagcctattc attgcttatg cttataagtt cctctccaat ggacaagagg 4860aaagaaaagg gtgactgtaa tcccgagctc atgagttcga ggctacagtc cgatggcagt 4920ggtgttgccg tctcgaacct agccgttaca cccttgtgca ttgtgg 496610640PRTBocavirus 10Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Thr Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Asp Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asn Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Cys Asp Pro Gly Arg His625 630 635 64011215PRTBocavirus 11Met Ser Ser Glu Ser Met Lys His Arg His Arg Ala Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Arg Arg Glu Arg Arg His Gln Trp Glu Asn His Lys 20 25 30Lys Ser Arg Arg Arg Ser Pro Ile Arg Gln His Gly Glu Lys Ser Leu 35 40 45Glu Ser Thr Pro Arg Cys Asn Gln Glu Ser Arg Gln Ser Ser Tyr Thr 50 55 60Ala Ser Lys Thr Ser Asp Gln Ala Thr Lys Thr Lys Glu Lys Thr Ser65 70 75 80Gly Gly Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala 85 90 95Asn His Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr 100 105 110Arg Leu Ala Arg Asp Gly Thr Asn Cys Ile Phe Asn Glu Met Lys Gln 115 120 125Glu Phe Gln Glu Leu Gln Ile Asn Gly Lys Ile Thr Trp Asp Asn Val 130 135 140Arg Glu Leu Leu Phe Ser Gln Lys Lys Lys Leu Asp Gln Lys Tyr Arg145 150 155 160Asn Met Leu Tyr His Phe Arg His Asn Ala Asp Cys Pro Arg Cys Asp 165 170 175Tyr Trp Asp Asp Val Tyr Arg Lys His Leu Ala His Val Ser Ser Gln 180 185 190Glu Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser 195 200 205Met Glu Thr Asn Ala Ser Asn 210 21512668PRTBocavirus 12Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Leu Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Pro Lys Asn Asn Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Gly Ser Met 130 135 140Asp Glu Gln Arg Gly Gly Gly Gly Gly Ala Val Gly Ser Val Gly Gly145 150 155 160Gly Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly 165 170 175Ser Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe 180 185 190Leu Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile 195 200 205Pro Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp 210 215 220Ser Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp225 230 235 240Trp Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met 245 250 255His Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly 260 265 270Ala Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe 275 280 285Cys Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu 290 295 300Asp Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr305 310 315 320Gly Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn 325 330 335Ala Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu 340 345 350Asn Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr 355 360 365Phe Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro 370 375 380Pro Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr385 390 395 400Ile Arg Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro 405 410 415Tyr Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala 420 425 430Gln Arg Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val 435 440 445Val Asn Pro Asp Gly Thr Ala Val Asn Ser Gly Met Ala Gly Val Gly 450 455 460Ser Gly Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu465 470 475 480Tyr Lys Ile Gln Trp Tyr Gln Thr Pro Glu Gly Thr Asn Ser Asp Gly 485 490 495Asn Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu 500 505 510Tyr Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met 515 520 525Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro 530 535 540Ile Trp Cys Lys Lys Pro Arg Ser Asp Lys Ser Thr Val Ile Asp Pro545 550 555 560Phe Asp Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile 565 570 575Lys Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr 580 585 590Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu 595 600 605Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr 610 615 620Ala Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Ile Asn Pro Thr Tyr625 630 635 640His Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met 645 650 655Cys Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 660 665134958DNABocavirus 13cgcacgtggt gagtgacact atggcctttt ctgctcctgt aattagagct ttttctcaac 60ctgctttcac ttatgttgtt aaatttccat atgataactg gaaagaggaa gagcacttac 120tatggagctt acttgctcct gggactgaac gtctcatgat ccaactaaga aactgcgcac 180cacatcctga agatgatcct gtcagggaag atattttatg ctcactagca gaccaacact 240atgctgctat tttcaccaaa gcttgctaca tggctgtaac ttcactcatg gggcagaaac 300agagaacaca ctttccacga tgcgacataa tttgccaggc tgagatcggc tcagaatatc 360tacactgtca catacttgtt ggaggagcag gtctgagcaa gagaaatgct aaaatttcat 420gtgctacgct cctaggcctt gtgatggctg aattaacaca acgctgcaaa ctacttcttg 480cacagcgtcc atttgaacca gatgaagcta gaatatttca tctactcaga cgcgttgaac 540gcgaagcatg gtcagggcac actggtaact gggttcaaat tcttcaatac agagacaagc 600gaggtgacct tcatgctcaa cacattgatc ctttacgctt tttcaaacac tacctgctgc 660caaaaaatcg gttgatctct ccttccagca agcctgacgt ctgcactact ccagataact 720ggtttgtcct agctgaaaaa acatacgctc acactattgt taatgggctt ccgctgctag 780aacataacag aaaggcctat ctacaagagt tagaaagtga agtcatccca gggccttcta 840ccatggcctt tgggggacgt ggtgcgtggg aacatctgcc tgaggtagga gaacaacgcc 900taataacttc taatacttct actgcttata aagctaacaa aaaagaaaaa ttaatgctaa 960acttacttga taaatgtgat gaacttaact tacttgtgta tgaagactta gttagtgctt 1020gtcctgacct tttacttatg cttgaaggtc agccaggtgg tgcacgccta attgaacagg 1080tgcttggcat gcatcatatt aaagtgtgtg ctaattacac agcgttatct ttcttgtttc 1140atctacatcc tgatcaatta ttaacttcta gcaataaagc actaaaacta ttgttgattc 1200aagggtacaa cccattgcaa gtaggccacg ccatctgttg tgttcttaac aaacagatgg 1260gcaagcagaa cacaatttgc ttttatggcc ctgcttcaac aggcaaaaca aatattgcaa 1320aggccatagt tcaaggcgtt cgtctgtatg gctgtgttaa tcatttaaac aaagggtttg 1380tctttaacga ttgcagacaa cgcctcataa tctggtggga ggagtgttta atgcaccaag 1440actgggtgga acctgctaaa tgcattctag gtggaactga atgtagaatt gatgttaaac 1500ataaggacag tgttcttctt caacaaacac cagtaattat ttccactaac catgacatct 1560actctgtagt tggtggcaat actgtatctc atgttcatgc agcgccctta aaagagcgaa 1620tccttcagct aaattttatg aaacaactgc cacaaacatt tggagaaatt tctccagttg 1680agattgcaga attactgcaa tggtgcttta atgagtacga ctgtactctt actggcttta 1740aacaaaaatg gaacttagat aaagttccaa actcatttcc tcttggggac ctttgtccta 1800cacattcaca ggactacgtg cttcacgaaa acggattctg cactgactgc ggcggctata 1860ttcctcatag tgctgacgac tctgtgtaca

ctgacgtggc tagcgaaaca tcaatcagca 1920gcgacgaccc aggtaggcat taatacatta gccttttaat atactacctt ccaagtgctt 1980atgtattaac tcctacaggt gacttggggg atacggacgg agagaactcc cagccggaga 2040catcgaacgt ggataatcgt ccatccaaga agagacgtgt gattccagaa actccaccaa 2100acagtccagt aagtcgccaa agcctttcta gctttttaga tacgtggcag tcacaaccta 2160gagacgaaga tgagctccga atctatgaag cacaggcatc gcgcatcaaa gagaacgccg 2220agtccactcc ggagagagag aagacaccag tgggagaacc acaagaagag tcgcagtcgg 2280agcccgatcc gacagcatgg ggagaaaagc ttggagtcta ctcctcgcta caaccaggag 2340agccgccaat cgtcttacac tgcttcgaag acctcagacc aagcgacgaa gacgaaggag 2400agaacatcgg gggggaatag aaccaatcct tatactgtgt tcagtcaaca cagggctaat 2460catccagatg ctcctggatg gtgtgggttt tactggcatt ctactcggct tgctagagat 2520ggcactaatt gtatctttaa tgaaatgaaa caagaatttc aagaactaca aataaatgga 2580aaaattactt gggataatgt tagagaattg ctgtttagcc agaaaaagaa actagatcag 2640aaatatagaa atatgctgta tcatttcaga cataacactg attgtcctag atgtgattat 2700tgggatgatg tataccgtaa acacttagct catgtctctt cacaggaatc agaggaggta 2760acagacgaag aaatgctttc tgctgttgaa agcatggaaa caaatgcctc caattaaacg 2820ccaacctgga gggtgggtgc ttcctggtta taaatacctt ggtccattta atcctcttga 2880aaacggtgaa ccagttaata aagctgatcg tgctgctcaa gctcatgata aatcatattc 2940tgaattaata aagagtggaa aaaaccctta cttatatttt aataaagctg atgaaaaatt 3000cattgacgat ttgaaaaacg actggtctgt tggtggcatt attggctcaa gtttctttaa 3060acttaagcgc gccgtggctc ctgctctagg aaataaagag cgagctcaaa aaagacattt 3120ttactttgca aactcaaata aaggtgctaa aagatcaaaa aacagcgaac ctaagccaag 3180cacttcaaaa atgtctgaaa atgaaattca agaccaacaa ccatcagact ctatggatgg 3240acaacgaggg ggcggaggag gtgcgactgg cagtgtggga ggggggaaag gttctggtgt 3300gggtatatcc acaggcggat gggtaggagg cagctacttt actgactcat atgtcataac 3360aaaaaacacc agacaatttc tagtaaagat acaaaatgac cacaaataca gaacagaaaa 3420tattattcca agcaatgctg gaggaaaatc acaaagatgc gtcagcacac cgtggtcata 3480ttttaacttc aatcaataca gcagtcactt ttcaccacaa gactggcagc gcctaacaaa 3540tgaatacaag cgctttaaac ctagaaaaat gcatgtaaaa atttacaacc tacaaataaa 3600acaaatactg tcaaatggtg ctgacactac atacaacaac gacctaacag ctggtgttca 3660catcttctgt gatggtgagc atgcgtatcc aaacgccaca cacccatggg acgaagatgt 3720aatgccagaa cttccatatg agacatggta tctgtttcaa tatggataca ttccagttat 3780tcatgaactt gctgaaatgg aagatgcaaa tgctgtagaa aaagctatag cactacaaat 3840accattcttc atgcttgaaa acagtgacca tgaagttcta agaactggag aaagcacaga 3900attcactttt gactttgact gtgaatggat caataacgaa agagcataca ttcctcctgg 3960attaatgttt aatccaaagg ttcctacaag aagagctcaa tacataagac aacacggaag 4020cacagcatca agcaacacca gaattccacc atatgcaaaa cctacaagct ggatgacagg 4080accaggtcta ctcagcgcac agagagtagg accagctgct tcagactcag cagcatggat 4140ggttgttgta aatccagacg gagctgctat taactcagga atggcaggaa ttggtacagg 4200ctttgatcct cctggtggat ctttaagacc aactgattta gaatataaaa tacaatggta 4260ccaaactcct gcaggtacaa acagcgatgg aaacatcatt tcaaatccat cattatccat 4320gcttagagat caagctctct acagaggaaa ccagacaaca tataacctat gttcagacgt 4380atggatgttc ccaaatcaaa tttgggacag atatccaata accagagaaa atccaatctg 4440gtgtaaaaaa ccaagatcag acaaaagcac aataattgat ccattcgatg gatcaattgc 4500aatggatcat cctccaggaa caattttcat aaaaatggca aaaattccag ttccttcaaa 4560caacaatgca gactcatact taaacatcta ctgcactgga caagtcagct gcgaaattgt 4620ctgggaagtt gaaagatatg caacaaagaa ctggagacca gaaagaagac acacggcact 4680tggccttggg attggaggag aagagaatgt aaatccaact taccatgttg acaaaaatgg 4740aaaatacatt cagccaacaa catgggacat gtgctatcct atcaaaacaa acatcaataa 4800agtgttgtaa ccttgtaagc ctcttttttg cttatgcttg taagttcctc tccaatggac 4860aagtggaaag aaaagggtga ctgtaatccc gagctcataa gttcgaggct acagtccgat 4920ggcagtggtg ttgccgtctc gaacctagcc gttacacc 495814640PRTBocavirus 14Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Asp Asp Pro Gly Arg His625 630 635 64015215PRTBocavirus 15Met Ser Ser Glu Ser Met Lys His Arg His Arg Ala Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Arg Arg Glu Arg Arg His Gln Trp Glu Asn His Lys 20 25 30Lys Ser Arg Ser Arg Ser Pro Ile Arg Gln His Gly Glu Lys Ser Leu 35 40 45Glu Ser Thr Pro Arg Tyr Asn Gln Glu Ser Arg Gln Ser Ser Tyr Thr 50 55 60Ala Ser Lys Thr Ser Asp Gln Ala Thr Lys Thr Lys Glu Arg Thr Ser65 70 75 80Gly Gly Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala 85 90 95Asn His Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr 100 105 110Arg Leu Ala Arg Asp Gly Thr Asn Cys Ile Phe Asn Glu Met Lys Gln 115 120 125Glu Phe Gln Glu Leu Gln Ile Asn Gly Lys Ile Thr Trp Asp Asn Val 130 135 140Arg Glu Leu Leu Phe Ser Gln Lys Lys Lys Leu Asp Gln Lys Tyr Arg145 150 155 160Asn Met Leu Tyr His Phe Arg His Asn Thr Asp Cys Pro Arg Cys Asp 165 170 175Tyr Trp Asp Asp Val Tyr Arg Lys His Leu Ala His Val Ser Ser Gln 180 185 190Glu Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser 195 200 205Met Glu Thr Asn Ala Ser Asn 210 21516668PRTBocavirus 16Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Val Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Arg Ser Lys Asn Ser Glu Pro Lys Pro Ser Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met 130 135 140Asp Gly Gln Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly145 150 155 160Gly Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly 165 170 175Ser Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe 180 185 190Leu Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile 195 200 205Pro Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp 210 215 220Ser Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp225 230 235 240Trp Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met 245 250 255His Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly 260 265 270Ala Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe 275 280 285Cys Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu 290 295 300Asp Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr305 310 315 320Gly Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn 325 330 335Ala Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu 340 345 350Asn Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr 355 360 365Phe Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro 370 375 380Pro Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr385 390 395 400Ile Arg Gln His Gly Ser Thr Ala Ser Ser Asn Thr Arg Ile Pro Pro 405 410 415Tyr Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala 420 425 430Gln Arg Val Gly Pro Ala Ala Ser Asp Ser Ala Ala Trp Met Val Val 435 440 445Val Asn Pro Asp Gly Ala Ala Ile Asn Ser Gly Met Ala Gly Ile Gly 450 455 460Thr Gly Phe Asp Pro Pro Gly Gly Ser Leu Arg Pro Thr Asp Leu Glu465 470 475 480Tyr Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly 485 490 495Asn Ile Ile Ser Asn Pro Ser Leu Ser Met Leu Arg Asp Gln Ala Leu 500 505 510Tyr Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met 515 520 525Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro 530 535 540Ile Trp Cys Lys Lys Pro Arg Ser Asp Lys Ser Thr Ile Ile Asp Pro545 550 555 560Phe Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile 565 570 575Lys Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr 580 585 590Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu 595 600 605Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr 610 615 620Ala Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr625 630 635 640His Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met 645 650 655Cys Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 660 665174948DNABocavirus 17gtgacactat ggccttttct gctcctgtaa ttagagcttt ttctcaacct gctttcactt 60atgttgttaa atttccatat gataactgga aagaggaaga gcacttacta tggagcttac 120ttgctcctgg gactgaacgt ctcatgatcc aactaagaaa ctgcgcacca catcctgaag 180atgatcctgt cagggaagat attttatgct cactagcaga ccaacactat gctgctattt 240tcaccaaagc ttgctacatg gctgtaactt cactcatggg gcagaaacag agaacacact 300ttccacgatg cgacataatt tgccaggctg agatcggctc agaatatcta cactgtcaca 360tacttgttgg aggagcaggt ctgagcaaga gaaatgctaa aatttcatgt gctacgctcc 420taggccttgt gatggctgaa ttaacacaac gctgcaaact acttcttgca cagcgtccat 480ttgaaccaga tgaagctaga atatttcatc tactcagacg cgttgaacgc gaagcatggt 540cagggcacac tggtaactgg gttcaaattc ttcaatacag agacaagcga ggtgaccttc 600atgctcaaca cattgatcct ttacgctttt tcaaacacta cctgctgcca aaaaatcggt 660tgatctctcc ttccagcaag cctgacgtct gcactactcc agataactgg tttgtcctag 720ctgaaaaaac atacgctcac actattgtta atgggcttcc gctgctagaa cataacagaa 780aggcctatct acaagagtta gaaagtgaag tcatcccagg gccttctacc atggcctttg 840ggggacgtgg tgcgtgggaa catctgcctg aggtaggaga acaacgccta ataacttcta 900atacttctac tgcttataaa gctaacaaaa aagaaaaatt aatgctaaac ttacttgata 960aatgtgatga acttaactta cttgtgtatg aagacttagt tagtgcttgt cctgaccttt 1020tacttatgct tgaaggtcag ccaggtggtg cacgcctaat tgaacaggtg cttggcatgc 1080atcatattaa agtgtgtgct aattacacag cgttatcttt cttatttcat ctacatcctg 1140atcaattatt aacttctagc aataaagcac taaaactatt gttgattcaa gggtacaacc 1200cattgcaagt aggccacgcc atctgttgtg tccttaacaa acagatgggc aaacagaaca 1260caatttgctt ttatggccct gcttcaacag gcaaaacaaa tattgcaaag gccatagttc 1320aaggcgttcg tctgtatggc tgtgttaatc atttaaacaa agggtttgtc tttaacgatt 1380gcagacaacg cctcataatc tggtgggagg agtgtttaat gcaccaagac tgggtagaac 1440ctgctaaatg cattctaggt ggaactgaat gtagaattga tgttaaacat aaggacagtg 1500ttcttcttca acaaacacca gtaattattt ccactaacca tgacatctac tctgtagttg 1560gtggcaatac tgtatctcat gttcatgcag cgcccttaaa agagcgaatc cttcagctaa 1620attttatgaa acaactgcca caaacatttg gagaaatttc tccagttgaa attgcagaat 1680tactgcaatg gtgctttaat gagtacgact gtactcttac tggctttaaa caaaaatgga 1740acttagataa agttccaaac tcatttcctc ttggggacct ttgtcctaca cattcacagg 1800actacgtgct tcacgaaaac ggattctgca ctgactgcgg cggctatatt cctcatagtg 1860ctgacgactc tgtgtacact gacgtggcta gcgagacatc aatcagcagc gacgacccag 1920gtaggcatta atacattagc cttttaatat actaccttcc aagtgcttat gtattaactc 1980ctacaggtga cttgggggat acggacggag agaactccca gccggagaca tcgaacgtgg 2040ataatcgtcc atccaagaag agacgtgtga ttccagaaac tccaccaaac agtccagtaa 2100gtcgccaaag cctttctagc tttttagata cgtggcagtc acaacctaga gacgaagatg 2160agctccgaat ctatgaagca caggcatcgc gcatcaaaga gaacgccgag tccactccgg 2220agagagagaa gacaccagtg ggagaaccac aagaagagtc gcagtcggag cccgatccga 2280cagcatgggg agaaaagctt ggagtctact cctcgctaca accaggagag ccgccaatcg 2340tcttacactg cttcgaagac ctcagaccaa gcgacgaaga cgaaggagag aacatcgggg 2400gggaatagaa ccaatcctta tactgtgttc agtcaacaca gggctaatca tccagatgct 2460cctggatggt gcgggtttta ctggcattct actcggcttg ctagagatgg cactaattgt 2520atctttaatg aaatgaaaca agaatttcaa gaactacaaa taaatggaaa aattacttgg 2580gataatgtta gagaattgct gtttagccag aaaaagaaac tagatcagaa atatagaaat 2640atgctgtatc

atttcagaca taacactgat tgtcctagat gtgattattg ggatgatgta 2700taccgtaaac acttagctca tgtctcttca caggaatcag aggaggtaac agacgaagaa 2760atgctttctg ctgttgaaag catggaaaca aatgcctcca attaaacgcc aacctggagg 2820gtgggtgctt cctggttata aataccttgg tccatttaat cctcttgaaa acggtgaacc 2880agttaataaa gctgatcgtg ctgctcaagc tcatgataaa tcatattctg aattaataaa 2940gagtggaaaa aacccttact tatattttaa taaagctgat gaaaaattca ttgacgattt 3000gaaaaacgac tggtctgttg gtggcattat tggctcaagt ttctttaaac ttaagcgcgc 3060cgtggctcct gctctaggaa ataaagagcg agctcaaaaa agacattttt actttgcaaa 3120ctcaaataaa ggtgctaaaa aatcaaaaaa cagcgaacct aagccaggca cttcaaaaat 3180gtctgaaaat gaaattcaag accaacaacc atcagactct atggatggac aacgaggggg 3240cggaggaggt gcggctggca gtgtgggagg ggggaaaggt tctggtgtgg gtatatccac 3300aggcggatgg gtaggaggca gctactttac tgactcatat gtcataacaa aaaacaccag 3360acaatttcta gtaaaaatac aaaatgacca caaatacaga acagaaaata ttattccaag 3420caatgctgga ggaaaatcac aaagatgcgt cagcacaccg tggtcatatt ttaacttcaa 3480tcaatacagc agtcactttt caccacaaga ctggcagcgc ctaacaaatg aatacaagcg 3540ctttaagcct agaaaaatgc atgtaaaaat ttacaaccta caaataaaac aaatactgtc 3600aaatggtgct gacactacat acaacaacga cctaacagct ggtgttcaca tcttctgtga 3660tggtgagcat gcgtatccaa acgccacaca cccatgggac gaagatgtaa tgccagaact 3720tccatatgag acatggtatc tgtttcaata tggatacatt ccagttattc atgaacttgc 3780tgaaatggaa gatgcaaatg ctgtagaaaa agctatagca ctacaaatac cattcttcat 3840gcttgaaaac agtgaccatg aagttctaag aactggagaa agcacagaat tcacttttga 3900ctttgactgt gaatggatca acaacgaaag agcatacatt cctcctggat taatgtttaa 3960tccaaaggtt cctacaagaa gagctcaata cataagacaa cacggaagca cagcatcaag 4020caacaccaga attccaccat atgcaaaacc tacaagctgg atgacaggac caggtctact 4080cagcgcacag agagtaggac cagctgcttc agactcagca gcatggatgg ttgttgtaaa 4140tccagacgga gctgctatta actcaggaat ggcaggaatt ggtacaggct ttgatcctcc 4200tggtggatct ttaagaccaa ctgatttaga atataaaata caatggtacc aaactcctgc 4260aggtacaaac agcgatggaa acatcatttc aaatccatca ttatccatgc ttagagatca 4320agctctctac agaggaaacc agacaacata taacctatgt tcagacgtat ggatgttccc 4380aaatcaaatt tgggacagat atccaataac aagagaaaat ccaatctggt gtaaaaaacc 4440aagatcagac aaaagcacaa taattgatcc attcgatgga tcaattgcaa tggatcatcc 4500tccaggaaca attttcataa aaatggcaaa aattccagtt ccttcaaaca acaatgcaga 4560ctcatactta aacatatact gcactggaca agtcagctgc gaaattgtct gggaagttga 4620aagatatgca acaaagaact ggagaccaga aagaagacac acggcacttg gccttgggat 4680tggaggagaa gaaaatgtaa atccaactta ccatgttgac aaaaatggta aatacattca 4740gccaacaact tgggacatgt gctatcctat caaaacaaac atcaataaag tgttgtaatc 4800tcttaagcct gttcattgct tatgcttata agttcctctc caatggacaa gaggaaagaa 4860aagggtgact gtaatcccga gctcatgagt tcgaggctac agtccgatgg cagtggtgtt 4920gccgtctcga acctagccgt tacaccct 494818640PRTBocavirus 18Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Asp Asp Pro Gly Arg His625 630 635 64019215PRTBocavirus 19Met Ser Ser Glu Ser Met Lys His Arg His Arg Ala Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Arg Arg Glu Arg Arg His Gln Trp Glu Asn His Lys 20 25 30Lys Ser Arg Ser Arg Ser Pro Ile Arg Gln His Gly Glu Lys Ser Leu 35 40 45Glu Ser Thr Pro Arg Tyr Asn Gln Glu Ser Arg Gln Ser Ser Tyr Thr 50 55 60Ala Ser Lys Thr Ser Asp Gln Ala Thr Lys Thr Lys Glu Arg Thr Ser65 70 75 80Gly Gly Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala 85 90 95Asn His Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr 100 105 110Arg Leu Ala Arg Asp Gly Thr Asn Cys Ile Phe Asn Glu Met Lys Gln 115 120 125Glu Phe Gln Glu Leu Gln Ile Asn Gly Lys Ile Thr Trp Asp Asn Val 130 135 140Arg Glu Leu Leu Phe Ser Gln Lys Lys Lys Leu Asp Gln Lys Tyr Arg145 150 155 160Asn Met Leu Tyr His Phe Arg His Asn Thr Asp Cys Pro Arg Cys Asp 165 170 175Tyr Trp Asp Asp Val Tyr Arg Lys His Leu Ala His Val Ser Ser Gln 180 185 190Glu Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser 195 200 205Met Glu Thr Asn Ala Ser Asn 210 21520668PRTBocavirus 20Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Val Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Ser Lys Asn Ser Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met 130 135 140Asp Gly Gln Arg Gly Gly Gly Gly Gly Ala Ala Gly Ser Val Gly Gly145 150 155 160Gly Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly 165 170 175Ser Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe 180 185 190Leu Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile 195 200 205Pro Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp 210 215 220Ser Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp225 230 235 240Trp Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met 245 250 255His Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly 260 265 270Ala Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe 275 280 285Cys Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu 290 295 300Asp Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr305 310 315 320Gly Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn 325 330 335Ala Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu 340 345 350Asn Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr 355 360 365Phe Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro 370 375 380Pro Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr385 390 395 400Ile Arg Gln His Gly Ser Thr Ala Ser Ser Asn Thr Arg Ile Pro Pro 405 410 415Tyr Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala 420 425 430Gln Arg Val Gly Pro Ala Ala Ser Asp Ser Ala Ala Trp Met Val Val 435 440 445Val Asn Pro Asp Gly Ala Ala Ile Asn Ser Gly Met Ala Gly Ile Gly 450 455 460Thr Gly Phe Asp Pro Pro Gly Gly Ser Leu Arg Pro Thr Asp Leu Glu465 470 475 480Tyr Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly 485 490 495Asn Ile Ile Ser Asn Pro Ser Leu Ser Met Leu Arg Asp Gln Ala Leu 500 505 510Tyr Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met 515 520 525Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro 530 535 540Ile Trp Cys Lys Lys Pro Arg Ser Asp Lys Ser Thr Ile Ile Asp Pro545 550 555 560Phe Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile 565 570 575Lys Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr 580 585 590Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu 595 600 605Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr 610 615 620Ala Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr625 630 635 640His Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met 645 650 655Cys Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 660 665214948DNABocavirus 21tccacctgtt attagagcat tctcttcccc tgcttttact tatgtcttca aatttccata 60tcgatcatgg aaagaaaaag aatggcttct tcatgcgctt ctagctcatg gtaccgagca 120agccatgatc cagctgagaa actgtgttcc tcatccggat gaagatataa tccgtgatga 180cttactgctt tctctagaag atcgccattt tggggcaatt ctctgcaaag ctgtttatat 240ggctactact actttcatgt cacagaaaca aagaaatatg tttcctcgct gtgacataat 300agtccagtct gagcttgggg agacaaacct acactgccat attatagttg ggggagaagg 360cttaagcaag agaaatgcaa aaacatcatg tcctcaacta tatggactga tactagggga 420attaatccaa cgctgcaaaa ctcttctggc tacgcgtcct ttcgaaccgg aagaggcaga 480aatttatcat gctttaaaac gagctgagcg agaagcttgg ggtggagtta ctagcggcaa 540cctacaaatt ctccaataca gagatcgcag aggagacctt cacgcacaac aagtggatgc 600tcttcgcttc ttcaaaaact acctattgcc taaaaataga tgcattacat cttacagcag 660acctgatgtc tgtacttctc cagaaaactg gtttgtttta gctgaaaaaa cttactgtca 720cactcttgtt aacgggctgc cgcttccaga acattacaga aaacactacc acgcaaccct 780agataacgaa gttctaccgg ggcctcagac aatggccttt gggggacgtg gtccgtggga 840acatcttcct gaggtaggag atcaacgttt agctgcctct tctgttagta caacatataa 900accaaacaaa aaagaaaaac ttatgcttaa cttactagat aaatgcagcg aattaaacct 960tttagtttat gaagacttag tagctaactg tcctgaactt ttgcttatgc ttgaaggtca 1020accaggtgga gcacgcctaa tagaacaagt cctaggcatg caccatatta atgtttgctc 1080taactttact gctcttagtt atctctttca cctttaccct agcacaactt tatcttcaga 1140taacaaggct ttgcagctgt tgttgataca aggttacaac ccattaatgg ttggtcacgc 1200cttgtgctgt gtactcaaca agcagtttgg caaacagaac actgtttgct tttatggacc 1260agcttctact ggtaaaacaa acatggcaaa ggccatagtc caaggcattc gactatatgg 1320ctgtgttaat catttaaaca aagggtttgt ctttaatgat tgcagacaac gcctagttgt 1380ttggtgggag gagtgcttaa tgcaccagga ttgggtggaa ccagcaaagt gtatcttggg 1440tggaactgag tgtagaattg acgtcaaaca cagagatagt gtattattga cacaaactcc 1500agtaattatt tccactaacc acgatatcta cgcggttgtt ggtggtaatt ctgtttctca 1560tgttcatgcg gctccattaa aagaaagagt gattcagcta aattttatga aacaacttcc 1620tcaaaccttt ggagagatca ctccagaaga aattgcagct ctactgcaat ggtgtttcaa 1680tgagtacgag tgtactctga caggctttaa aacaaaatgg agcctagata agattccaaa 1740ctcatttcct cttggggtcc tttgtcctac tcattcacag gacttcatac tccacgaaaa 1800cggatactgc actgattgtg gtggttacct tgctcatagc gctgacgatt ctgtgtacac 1860tgatcgtgca agcgacacta gcaaagaagc catcgacgca ggtaagttta cattctccag 1920acacttcata tatatcctac acaccacact aaaacatatg tttaattaca ggtgacttgg 1980gggatacgga cggagaggac tccgagtctg aagcatcgga agtgggtgtt cgtccatcca 2040agaagcgacg cataactatt cctgcaactc caccaaattc tcctggcagc tctgtgagta 2100cttctgcctt ctttgataat tggtgcgcac aaccgcgaga cgaagatgag ctcagggaat 2160acaaaagaca agcatcgcgc ctacaaaaga aaagggagtc cagggagaga cgagaggaaa 2220cgcccatggc aacctcgtca caggagtcgg agccggagcc caatccgacg cagtgggaag 2280acaagctcgg ggtcataccg tcaggaacac cagatcagcc acctatcgtc ttgcactgct 2340tcgaagatct cagacccagt gacgaagacg aaggagaata catcggggaa gagagactct 2400agaactaatc catatactgt attcagccag cataaagctt caaatcctga tgctccaggg 2460tggtgtgggt tttattggca ctctactaga attgctagaa atggtactaa tgcaatcttt 2520aatgaaatga aacagcagtt tcaacaactg caactagaca acaagattgg ctgggataat 2580gctagagaac tattgtttag ccagaaaaaa tcactagatc aacaatacag aaatatgttt 2640tggcacttta gaaatgctcc tgattgtgaa cgctgtaatt actgggataa tgtgtaccgt 2700atgcacttag ctcatgtttc ctctcaggca gaatcagaag aaataactga cgaggaaatg 2760ctttctgctg ctgaaagtat ggaaacagat gcctccaatt aaaaggcaac ctggagggtg 2820ggtacttcct ggttacaaat accttggtcc atttaatcct cttgataacg gtgaaccagt 2880taataaagct gatcgtgctg ctcaagctca tgataaatca tattctgaat taataaaaaa 2940gtggaaaaaa atccttactt gtattttcaa taaagctgat gaaaaattca ttgacgattt 3000gaaaaacgac tggtctcttg gtggcattat tggctcaaat ttctttaaac ttaagcgcgc 3060cgtggctcct gctctaggaa ataaagagcg agctcaaaaa agacattttt attttgcaaa 3120ctcaaataaa ggtgctaaaa aatcaaaaaa caacgaacct aaaccaagca cctcaaaaat 3180gtctgaaaat gaaattcaag accaacagcc atcagaacct aatgatggcc agcgaggagg 3240gggcggaggt acaaccggca gtgtgggagg ggggaaaggt tctggtgtgg gtatatccac 3300aggtggatgg gtaggaggca gctactttac tgactcatat gtcataacaa aaaacaccag 3360acaatttctg gttaaaatcc aaaacaacca

tcaatataaa actgaaaata taattccttc 3420taatggagga ggaaaatcac aaagatgtgt cagcacacca tggtcatact ttaactttaa 3480tcaatacagc agccatttct caccacaaga ctggcagcgc ctaacaaatg aatacaaaag 3540attcagacct aaaggtatgc atgttaaaat ctacaattta cagataaaac aaattttatc 3600aaatggtgct gatgttacat acaacaacga cctaacagca ggagtacaca tcttttgtga 3660tggcgaacat gcatatccaa acgctacaca tccatgggac gaagatgtga tgccagagct 3720tccttaccaa acatggtatc tatttcaata tggatacata cctacaattc atgaacttgc 3780agaaatggaa gactccaatg cagtagaaaa agcaattgct ttacaaatac cattcttcat 3840gcttgaaaac agcgaccatg aagttctaag aactggagaa agtgcagaat tcaactttaa 3900ctttgattgt gaatggatta acaatgaaag agcattcatt cctccaggac tgatgttcaa 3960tccattggta ccaacaagaa gagctcaata cataagaaga aatggaaaca ctcaagcaag 4020taccacaaga atccaacctt actcaaaacc aacaagttgg atgactggac caggtctact 4080cagtgcgcaa cgagtaggtc cagctgcttc agacacagct gcatggatgg ttggaataga 4140tctagacggt gcaaacgtaa actcaggaag agcaggagtc agcacaggct ttgatcctcc 4200agcaggttca cttagaccta cagatctaga atacaaaata caatggtacc agactccagc 4260tggaacaaac aatgatggaa acatcatatc aaatccacct ctttcaatgc tcagagatca 4320aactctatac aaaggaaacc aaacaaccta caacttatgc tcagatgtat ggatgttccc 4380aaatcaaatt tgggacagat acccaataac cagagaaaat cctatttggt gcaaacaacc 4440aagatcagac aaacacactg tcattgatcc gttcgatgga tctcttgcaa tggatcatcc 4500tccaggaaca atttttatca aaatggcaaa aattccagtt ccttcaagca acaacgcaga 4560ctcatactta aacatctact gcactggaca agtcagctgc gaaattgtct gggaagtcga 4620aagatatgca acaaagaact ggagaccaga aagaagacac acggcactcg gccttggaat 4680tggaggagca gatgaaatca acccaacata ccatgttgac aaaaacggag cctacattca 4740accaacatca tgggacatgt gctttccagt taaaacaaac atcaataaag tgttgtaatc 4800tcttaagcct ctttattgct cacgcttgta agttcctctc caatggacaa gtggaaagaa 4860aagggtgact gtaatcccga gctcatgagt tcgaggctac agtccgatgg cagtggtgtt 4920gccgtctcga acctagccgt taaaccct 494822657PRTBocavirus 22Pro Pro Val Ile Arg Ala Phe Ser Ser Pro Ala Phe Thr Tyr Val Phe1 5 10 15Lys Phe Pro Tyr Arg Ser Trp Lys Glu Lys Glu Trp Leu Leu His Ala 20 25 30Leu Leu Ala His Gly Thr Glu Gln Ala Met Ile Gln Leu Arg Asn Cys 35 40 45Val Pro His Pro Asp Glu Asp Ile Ile Arg Asp Asp Leu Leu Leu Ser 50 55 60Leu Glu Asp Arg His Phe Gly Ala Ile Leu Cys Lys Ala Val Tyr Met65 70 75 80Ala Thr Thr Thr Phe Met Ser Gln Lys Gln Arg Asn Met Phe Pro Arg 85 90 95Cys Asp Ile Ile Val Gln Ser Glu Leu Gly Glu Thr Asn Leu His Cys 100 105 110His Ile Ile Val Gly Gly Glu Gly Leu Ser Lys Arg Asn Ala Lys Thr 115 120 125Ser Cys Pro Gln Leu Tyr Gly Leu Ile Leu Gly Glu Leu Ile Gln Arg 130 135 140Cys Lys Thr Leu Leu Ala Thr Arg Pro Phe Glu Pro Glu Glu Ala Glu145 150 155 160Ile Tyr His Ala Leu Lys Arg Ala Glu Arg Glu Ala Trp Gly Gly Val 165 170 175Thr Ser Gly Asn Leu Gln Ile Leu Gln Tyr Arg Asp Arg Arg Gly Asp 180 185 190Leu His Ala Gln Gln Val Asp Ala Leu Arg Phe Phe Lys Asn Tyr Leu 195 200 205Leu Pro Lys Asn Arg Cys Ile Thr Ser Tyr Ser Arg Pro Asp Val Cys 210 215 220Thr Ser Pro Glu Asn Trp Phe Val Leu Ala Glu Lys Thr Tyr Cys His225 230 235 240Thr Leu Val Asn Gly Leu Pro Leu Pro Glu His Tyr Arg Lys His Tyr 245 250 255His Ala Thr Leu Asp Asn Glu Val Leu Pro Gly Pro Gln Thr Met Ala 260 265 270Phe Gly Gly Arg Gly Pro Trp Glu His Leu Pro Glu Val Gly Asp Gln 275 280 285Arg Leu Ala Ala Ser Ser Val Ser Thr Thr Tyr Lys Pro Asn Lys Lys 290 295 300Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Ser Glu Leu Asn Leu305 310 315 320Leu Val Tyr Glu Asp Leu Val Ala Asn Cys Pro Glu Leu Leu Leu Met 325 330 335Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu Gln Val Leu Gly 340 345 350Met His His Ile Asn Val Cys Ser Asn Phe Thr Ala Leu Ser Tyr Leu 355 360 365Phe His Leu Tyr Pro Ser Thr Thr Leu Ser Ser Asp Asn Lys Ala Leu 370 375 380Gln Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Met Val Gly His Ala385 390 395 400Leu Cys Cys Val Leu Asn Lys Gln Phe Gly Lys Gln Asn Thr Val Cys 405 410 415Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Met Ala Lys Ala Ile 420 425 430Val Gln Gly Ile Arg Leu Tyr Gly Cys Val Asn His Leu Asn Lys Gly 435 440 445Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Val Val Trp Trp Glu Glu 450 455 460Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys Cys Ile Leu Gly465 470 475 480Gly Thr Glu Cys Arg Ile Asp Val Lys His Arg Asp Ser Val Leu Leu 485 490 495Thr Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp Ile Tyr Ala Val 500 505 510Val Gly Gly Asn Ser Val Ser His Val His Ala Ala Pro Leu Lys Glu 515 520 525Arg Val Ile Gln Leu Asn Phe Met Lys Gln Leu Pro Gln Thr Phe Gly 530 535 540Glu Ile Thr Pro Glu Glu Ile Ala Ala Leu Leu Gln Trp Cys Phe Asn545 550 555 560Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Thr Lys Trp Ser Leu Asp 565 570 575Lys Ile Pro Asn Ser Phe Pro Leu Gly Val Leu Cys Pro Thr His Ser 580 585 590Gln Asp Phe Ile Leu His Glu Asn Gly Tyr Cys Thr Asp Cys Gly Gly 595 600 605Tyr Leu Ala His Ser Ala Asp Asp Ser Val Tyr Thr Asp Arg Ala Ser 610 615 620Asp Thr Ser Lys Glu Ala Ile Asp Ala Gly Lys Phe Thr Phe Ser Arg625 630 635 640His Phe Ile Tyr Ile Leu His Thr Thr Leu Lys His Met Phe Asn Tyr 645 650 655Arg23218PRTBocavirus 23Met Ser Ser Gly Asn Thr Lys Asp Lys His Arg Ala Tyr Lys Arg Lys1 5 10 15Gly Ser Pro Gly Arg Asp Glu Arg Lys Arg Pro Trp Gln Pro Arg His 20 25 30Arg Ser Arg Ser Arg Ser Pro Ile Arg Arg Ser Gly Lys Thr Ser Ser 35 40 45Gly Ser Tyr Arg Gln Glu His Gln Ile Ser His Leu Ser Ser Cys Thr 50 55 60Ala Ser Lys Ile Ser Asp Pro Val Thr Lys Thr Lys Glu Asn Thr Ser65 70 75 80Gly Lys Arg Asp Ser Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His 85 90 95Lys Ala Ser Asn Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His 100 105 110Ser Thr Arg Ile Ala Arg Asn Gly Thr Asn Ala Ile Phe Asn Glu Met 115 120 125Lys Gln Gln Phe Gln Gln Leu Gln Leu Asp Asn Lys Ile Gly Trp Asp 130 135 140Asn Ala Arg Glu Leu Leu Phe Ser Gln Lys Lys Ser Leu Asp Gln Gln145 150 155 160Tyr Arg Asn Met Phe Trp His Phe Arg Asn Ala Pro Asp Cys Glu Arg 165 170 175Cys Asn Tyr Trp Asp Asn Val Tyr Arg Met His Leu Ala His Val Ser 180 185 190Ser Gln Ala Glu Ser Glu Glu Ile Thr Asp Glu Glu Met Leu Ser Ala 195 200 205Ala Glu Ser Met Glu Thr Asp Ala Ser Asn 210 21524669PRTBocavirus 24Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Asp Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Lys Trp Lys Lys Ile Leu Thr Cys Ile Phe Asn Lys Ala Asp 50 55 60Glu Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Leu Gly Gly Ile65 70 75 80Ile Gly Ser Asn Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu 85 90 95Gly Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser 100 105 110Asn Lys Gly Ala Lys Lys Ser Lys Asn Asn Glu Pro Lys Pro Ser Thr 115 120 125Ser Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Glu Pro 130 135 140Asn Asp Gly Gln Arg Gly Gly Gly Gly Gly Thr Thr Gly Ser Val Gly145 150 155 160Gly Gly Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly 165 170 175Gly Ser Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln 180 185 190Phe Leu Val Lys Ile Gln Asn Asn His Gln Tyr Lys Thr Glu Asn Ile 195 200 205Ile Pro Ser Asn Gly Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro 210 215 220Trp Ser Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln225 230 235 240Asp Trp Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Gly 245 250 255Met His Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn 260 265 270Gly Ala Asp Val Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile 275 280 285Phe Cys Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp 290 295 300Glu Asp Val Met Pro Glu Leu Pro Tyr Gln Thr Trp Tyr Leu Phe Gln305 310 315 320Tyr Gly Tyr Ile Pro Thr Ile His Glu Leu Ala Glu Met Glu Asp Ser 325 330 335Asn Ala Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu 340 345 350Glu Asn Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Ala Glu Phe 355 360 365Asn Phe Asn Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Phe Ile 370 375 380Pro Pro Gly Leu Met Phe Asn Pro Leu Val Pro Thr Arg Arg Ala Gln385 390 395 400Tyr Ile Arg Arg Asn Gly Asn Thr Gln Ala Ser Thr Thr Arg Ile Gln 405 410 415Pro Tyr Ser Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser 420 425 430Ala Gln Arg Val Gly Pro Ala Ala Ser Asp Thr Ala Ala Trp Met Val 435 440 445Gly Ile Asp Leu Asp Gly Ala Asn Val Asn Ser Gly Arg Ala Gly Val 450 455 460Ser Thr Gly Phe Asp Pro Pro Ala Gly Ser Leu Arg Pro Thr Asp Leu465 470 475 480Glu Tyr Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Asn Asp 485 490 495Gly Asn Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Thr 500 505 510Leu Tyr Lys Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp 515 520 525Met Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn 530 535 540Pro Ile Trp Cys Lys Gln Pro Arg Ser Asp Lys His Thr Val Ile Asp545 550 555 560Pro Phe Asp Gly Ser Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe 565 570 575Ile Lys Met Ala Lys Ile Pro Val Pro Ser Ser Asn Asn Ala Asp Ser 580 585 590Tyr Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp 595 600 605Glu Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His 610 615 620Thr Ala Leu Gly Leu Gly Ile Gly Gly Ala Asp Glu Ile Asn Pro Thr625 630 635 640Tyr His Val Asp Lys Asn Gly Ala Tyr Ile Gln Pro Thr Ser Trp Asp 645 650 655Met Cys Phe Pro Val Lys Thr Asn Ile Asn Lys Val Leu 660 665254946DNABocavirus 25actatggcct tttctgctcc tgtacttaga gctttttctc aacctacttt tacctatgtt 60attaaatttc catataataa ctggaaagaa gatgaacact tactatggag cttacttgct 120cctgggactg aaagtctcat gattcaacta aaaaactgcg caccacatcc tgaagatgat 180cctatcaggg aagatatttt atgctcacta gcagatctac actatggtgc tgtttttgcc 240aaagcttgct acatagctac atctacacta atggggcaga aacaaagaac actctttcca 300cgctgcgaca ttgtttgcca gtctgaaatt ggctcagact ttctacactg tcacatactt 360gttggaggag ccggtcttag caagagaaat gctaaaattt cacgcgctac acttttgggt 420cttgtgatgg ctgaactaac acaacgctgc aagctacttc ttgcacatcg tccatttgaa 480ccagctgaag ctactatcta tcatgaactt aaacgcattg aacgcgaagc atggtcaggg 540catactggta actgggttca gattcttcaa tacaaagata aacgaggtga tcttcacgct 600caaccaattg atcccttacg ctttctaaaa cattacattc taccaaaaaa tcgattgatt 660tctccttcca gcaaacctga cgtctgcact tctccagata actggtttat tctagctgac 720aaaacatact ctcacactat tattaatggg cttccgctgc tagaacgtaa cagaaaagcc 780tatctacaag agttagaaag tgaagtcatc ccggggcctt ctgccatggc ctttggggga 840cgtggtgcgt gggaacaact tcctgaggta ggagaacaac gcctaattac ttctaatact 900tctactgctt ataaagctaa caaaaaagaa aaattaatgt taaatttact tgataaatgt 960gatgaactta atttgcttgt atatgaagac ttagttagtg cttgtcctga ccttttacta 1020atgcttgaag gacaaccggg tggagcacga ctaattgaac aggtgcttgg catgcaccat 1080attaaagtat gtgctaaaca tactgcctta tcttttttat ttcacttaca tcctgatcaa 1140ttattaactt ctagcaataa agcactaaaa ctactattga ttcaaggata caacccatta 1200caagtagggc atgccatctg ttgtgtactt aacaaacaga tgggcaagca gaacactatt 1260tgcttttatg gccctgcttc aacaggcaaa acaaactttg caaaagctat agttcagggc 1320gttcgccttt atggctgtgt taatcattta aacaaggggt ttgtttttaa cgattgcaga 1380caacgcctta taatttggtg ggaagaatgt ttaatgcatc aagattgggt agaacctgct 1440aaatgtattt taggcggaac tgaatgtaga attgatgtta aacataaaga tagtgttctc 1500cttcaacaaa caccagtaat catttccact aaccatgaca tctactctgt agttggtggc 1560aatactgttt ctcatgttca tgcagcacca ttaaaagaac gagttcttca gctaaatttt 1620atgaaacaac taccacaaac atttggagaa atctctccaa gtgaaattgc agaacttttg 1680caatggtgct ttaatgagta cgactgtact cttgctggct ttaaacaaaa atggaactta 1740gacaaagttc caaactcatt tcctattgga gacctttgtc ctacacattc acaggacttc 1800acgcttcacg aaaacggatt ctgctctgac tgtggcggct atcttcctca tagcgctgac 1860gattctgttt acactgacgt ggctagtgaa acaaccagcg gtgactacga cccaggtagg 1920ctttaataca ttagctttac tatttattac tcttgaagtt tgcttatgta ttaactccta 1980caggtaacct gggggatacg gacggagagg actccaagtc agaagcatcg gaagtggatt 2040attgtccacc caagaaaagg cgtgtgattt cagcaactcc accaaacagt ccagtaagtg 2100gtccaagcct ttctaccttt ttagatactt ggcaatcaca acctagagac gacgatgagc 2160tcagaatcta cgaagaacag gcatcgcagt tccaaaagaa caccaagtcc acttcagaaa 2220gagaggaagc gcaactggga gaatcgcaag agccgcagcc ggagcccgat ccgacggcat 2280ggggagaaaa acttggagta tgctcatcac aacaaccagg acaaccgcca atcgtcctat 2340actgcttcga agacctcaga ccaagcgatg aagacgaagg agaaaacatc gggggggact 2400agaacaaatc cttatactgt attcagtcaa cacagggcta atcattcaaa tgctcctggc 2460tggtgtgggt tttactggca ttcaactagg cttgctagaa atggcactaa taatattttt 2520aatgaaatga aacaaaaatt tcaagaacta caaatagatg ggaaaatcag ttgggatact 2580actagagaac tattgtttac tcagaaaaaa acattagatc aaggctacag aaacatgttg 2640taccacttta gacacagtcc tgattgtcct agatgtgatt attgggatga tgtttaccgt 2700aaacacttag ctaatgtctc ttcacaggaa tcagaggagg ttacagacga agaaatgctt 2760tctgctgttg aaagcatgga aacaaatgcc tccaattaaa cgccaacctg gagggtgggt 2820gcttcctggt tataaatacc ttggtccatt taatcctctt gaaaacggtg aaccagttaa 2880taaagctgat cgtgctgctc aagctcatga taaatcatat tctgaactaa taaaaagtgg 2940aaaaaatcct tacttatatt tcaataaagc tgatgaaaaa ttcattgacg atttgaaaga 3000cgattggtct cttggtggca ttattggctc aagttttttt aaacttaaac gcgccgtggc 3060tcctgctcta ggaaataaag agcgagctca aaaaagacat ttctactttg caaactcaaa 3120taaaggtgct aaaaaaacaa aaaacaacga acctaagcca ggcacttcaa aaatgtctga 3180aaatgaaatt caagaccaac aaccatcaga ttctatggat ggacaacgag ggggcggagg 3240aggtgcaact ggcagtgtgg gaggggggaa aggttctggt gtgggtatat ccacaggcgg 3300atgggtagga ggcagctatt ttactgactc atatgttata acaaaaaaca ccagacaatt 3360tctagttaaa atacaaaaca accatcaata caaaacagaa ttaatatcgc cttccacatc 3420tcaaggaaaa tcacaaagat gcgtcagcac gccttggtct tactttaact ttaatcaata 3480cagcagtcat ttttcaccac aagactggca gcgattaaca aacgaatata aaagattcag 3540acccaaaggc atgcatgtta aaatatacaa tttacaaata aaacaaattc tttcaaatgg 3600tgctgacact acatacaaca acgacctcac agctggtgtt cacatttttt gtgatggcga 3660acacgcatat ccaaatgcaa cacatccttg ggatgaagac gttatgccag agctgccata 3720ccaaacatgg tatttgtttc aatatggata tattccagtt atacatgaac ttgctgaaat 3780ggaagactca aatgctgtag aaaaagcaat ttgcttacaa ataccatttt ttatgcttga 3840aaacagcgac cacgaagttt taagaacagg tgaaagcaca gaatttactt tcaactttga 3900ctgtgaatgg ataaacaatg aaagagcata cattcctcca ggcttaatgt ttaatccact 3960agtacctact agaagagcac agtacataag aagaaacaac aatcctcaaa ctgctgaaag 4020cacatccaga

attgctccat atgcaaaacc tacaagttgg atgactggac caggtttact 4080cagtgcacaa agagtaggtc cagctacttc agacacagga gcctggatgg ttgcagttaa 4140accagaaaac gcaagcattg acacaggaat gtctggaatt ggaagtggat ttgatccacc 4200acaaggatca ctagcaccaa caaatctaga atacaaaatc caatggtacc aaacaccaca 4260aggaacaaac aacaatggaa acatcatatc taatcaacca ctatctatgc taagagatca 4320agctttattt agaggaaatc aaacaaccta taacctatgt tcagatgtat ggatgtttcc 4380aaatcaaatt tgggacagat acccaataac cagagaaaat ccaatatggt gtaaaaaacc 4440cagatcagac aaacacacaa caattgatcc ttttgatgga tcccttgcaa tggatcatcc 4500tccaggcaca atttttatta aaatggcaaa aattccagtt ccttcaaaca acaatgcaga 4560ctcatactta aacatttact gcacagggca agtcagctgt gaaattgtct gggaagttga 4620aagatatgca acaaagaact ggagaccaga aagaagacac acaacatttg gtcttggaat 4680tggaggagct gacaacttaa atccaaccta ccatgttgac aaaaacggaa cttacattca 4740accaacaaca tgggacatgt gctttccagt taaaacaaac atcaataaag tgttgtaacc 4800ttctaagcct cttttttgct tatgcttata agttcctctc caatggacaa gtggaaagaa 4860aagggtgact gtaatcccga gctcatgagt tcgaggctac agtccgatgg cagtggtgtt 4920gccgtctcga acctagccgt tacacc 494626640PRTBocavirus 26Met Ala Phe Ser Ala Pro Val Leu Arg Ala Phe Ser Gln Pro Thr Phe1 5 10 15Thr Tyr Val Ile Lys Phe Pro Tyr Asn Asn Trp Lys Glu Asp Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Ser Leu Met Ile Gln 35 40 45Leu Lys Asn Cys Ala Pro His Pro Glu Asp Asp Pro Ile Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Leu His Tyr Gly Ala Val Phe Ala Lys65 70 75 80Ala Cys Tyr Ile Ala Thr Ser Thr Leu Met Gly Gln Lys Gln Arg Thr 85 90 95Leu Phe Pro Arg Cys Asp Ile Val Cys Gln Ser Glu Ile Gly Ser Asp 100 105 110Phe Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Arg Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala His Arg Pro Phe Glu Pro145 150 155 160Ala Glu Ala Thr Ile Tyr His Glu Leu Lys Arg Ile Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Lys Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln Pro Ile Asp Pro Leu Arg Phe Leu 195 200 205Lys His Tyr Ile Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Ser Pro Asp Asn Trp Phe Ile Leu Ala Asp Lys225 230 235 240Thr Tyr Ser His Thr Ile Ile Asn Gly Leu Pro Leu Leu Glu Arg Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Ala Met Ala Phe Gly Gly Arg Gly Ala Trp Glu Gln Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Lys His Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Phe 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Ser Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Ala Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Ile Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Phe Thr Leu His Glu Asn Gly Phe Cys Ser 595 600 605Asp Cys Gly Gly Tyr Leu Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Thr Ser Gly Asp Tyr Asp Pro Gly Arg Leu625 630 635 64027214PRTBocavirus 27Met Ser Ser Glu Ser Thr Lys Asn Arg His Arg Ser Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Gln Lys Glu Arg Lys Arg Asn Trp Glu Asn Arg Lys 20 25 30Ser Arg Ser Arg Ser Pro Ile Arg Arg His Gly Glu Lys Asn Leu Glu 35 40 45Tyr Ala His His Asn Asn Gln Asp Asn Arg Gln Ser Ser Tyr Thr Ala 50 55 60Ser Lys Thr Ser Asp Gln Ala Met Lys Thr Lys Glu Lys Thr Ser Gly65 70 75 80Gly Thr Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala Asn 85 90 95His Ser Asn Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr Arg 100 105 110Leu Ala Arg Asn Gly Thr Asn Asn Ile Phe Asn Glu Met Lys Gln Lys 115 120 125Phe Gln Glu Leu Gln Ile Asp Gly Lys Ile Ser Trp Asp Thr Thr Arg 130 135 140Glu Leu Leu Phe Thr Gln Lys Lys Thr Leu Asp Gln Gly Tyr Arg Asn145 150 155 160Met Leu Tyr His Phe Arg His Ser Pro Asp Cys Pro Arg Cys Asp Tyr 165 170 175Trp Asp Asp Val Tyr Arg Lys His Leu Ala Asn Val Ser Ser Gln Glu 180 185 190Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser Met 195 200 205Glu Thr Asn Ala Ser Asn 21028670PRTBocavirus 28Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asp Asp Trp Ser Leu Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Thr Lys Asn Asn Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met 130 135 140Asp Gly Gln Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly145 150 155 160Gly Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly 165 170 175Ser Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe 180 185 190Leu Val Lys Ile Gln Asn Asn His Gln Tyr Lys Thr Glu Leu Ile Ser 195 200 205Pro Ser Thr Ser Gln Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp 210 215 220Ser Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp225 230 235 240Trp Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Gly Met 245 250 255His Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly 260 265 270Ala Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe 275 280 285Cys Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu 290 295 300Asp Val Met Pro Glu Leu Pro Tyr Gln Thr Trp Tyr Leu Phe Gln Tyr305 310 315 320Gly Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ser Asn 325 330 335Ala Val Glu Lys Ala Ile Cys Leu Gln Ile Pro Phe Phe Met Leu Glu 340 345 350Asn Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr 355 360 365Phe Asn Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro 370 375 380Pro Gly Leu Met Phe Asn Pro Leu Val Pro Thr Arg Arg Ala Gln Tyr385 390 395 400Ile Arg Arg Asn Asn Asn Pro Gln Thr Ala Glu Ser Thr Ser Arg Ile 405 410 415Ala Pro Tyr Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu 420 425 430Ser Ala Gln Arg Val Gly Pro Ala Thr Ser Asp Thr Gly Ala Trp Met 435 440 445Val Ala Val Lys Pro Glu Asn Ala Ser Ile Asp Thr Gly Met Ser Gly 450 455 460Ile Gly Ser Gly Phe Asp Pro Pro Gln Gly Ser Leu Ala Pro Thr Asn465 470 475 480Leu Glu Tyr Lys Ile Gln Trp Tyr Gln Thr Pro Gln Gly Thr Asn Asn 485 490 495Asn Gly Asn Ile Ile Ser Asn Gln Pro Leu Ser Met Leu Arg Asp Gln 500 505 510Ala Leu Phe Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val 515 520 525Trp Met Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu 530 535 540Asn Pro Ile Trp Cys Lys Lys Pro Arg Ser Asp Lys His Thr Thr Ile545 550 555 560Asp Pro Phe Asp Gly Ser Leu Ala Met Asp His Pro Pro Gly Thr Ile 565 570 575Phe Ile Lys Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp 580 585 590Ser Tyr Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val 595 600 605Trp Glu Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg 610 615 620His Thr Thr Phe Gly Leu Gly Ile Gly Gly Ala Asp Asn Leu Asn Pro625 630 635 640Thr Tyr His Val Asp Lys Asn Gly Thr Tyr Ile Gln Pro Thr Thr Trp 645 650 655Asp Met Cys Phe Pro Val Lys Thr Asn Ile Asn Lys Val Leu 660 665 67029776PRTBocavirus 29Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Glu Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Gly Ala Ile Phe Ala Lys65 70 75 80Ala Cys Tyr Ile Ala Thr Thr Thr Leu Met Gly Gln Lys Gln Arg Thr 85 90 95Pro Phe Pro Arg Cys Asp Ile Ile Cys Gln Ser Glu Ile Gly Ser Glu 100 105 110His Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Arg Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Gln Leu Leu Ala Leu Arg Pro Phe Glu Pro145 150 155 160Ala Glu Ala Asn Ile Phe His Leu Leu Lys Arg Ile Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Lys Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln Pro Ile Asp Pro Leu Arg Phe Leu 195 200 205Lys His Tyr Ile Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Ile Leu Ala Asp Lys225 230 235 240Thr Tyr Ala His Thr Ile Ile Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu Gln Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Ala Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Lys Tyr Thr Ala 355 360 365Leu Thr Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Asn 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Phe 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Thr Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Leu Pro His Ser Ala Asp Asp Phe Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Thr Ser Gly Asp Cys Asp Pro Gly Asn Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asp Ser Lys Ser Glu Ala Ser Glu Val Asp 645 650 655Phe Arg Pro Ser Lys Lys Arg Arg Val Ile Ser Ala Thr Pro Pro Ser 660 665 670Ser Pro Val Ser Gly Pro Ser Leu Ser Thr Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Glu Asp Glu Leu Arg Ile Tyr Glu Glu Gln Ala 690 695 700Ser Gln Leu Gln Lys Asn Thr Lys Ser Thr Pro Glu Arg Glu Glu Ala705 710 715 720Gln Leu Gly Glu Pro Gln Glu Pro Gln Pro Glu Pro Asp Pro Thr Ala 725 730 735Trp Gly Glu Lys Leu Gly Val Cys Ser Ser Gln Gln Pro Gly Glu Pro 740 745 750Pro Val Val Leu Tyr Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu Asp 755 760 765Glu Gly Glu Asn Ile Gly Gly Glu 770 77530538PRTBocavirus 30Met Ser Glu Asn Glu Ile Gln

Asp Gln Gln Pro Ser Gly Ser Met Glu1 5 10 15Glu Arg Gly Gly Gly Gly Gly Ala Val Gly Ser Val Gly Gly Gly Lys 20 25 30Gly Ser Ser Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser Tyr 35 40 45Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu Val 50 55 60Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro Ser65 70 75 80Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser Tyr 85 90 95Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp Gln 100 105 110His Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His Val 115 120 125Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala Asp 130 135 140Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys Asp145 150 155 160Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp Val 165 170 175Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly Tyr 180 185 190Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala Val 195 200 205Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn Ser 210 215 220Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe Asp225 230 235 240Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro Gly 245 250 255Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile Arg 260 265 270Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr Ala 275 280 285Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln Arg 290 295 300Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val Asn305 310 315 320Pro Asp Gly Ala Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser Gly 325 330 335Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr Lys 340 345 350Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly Asn Ile 355 360 365Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr Arg 370 375 380Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe Pro385 390 395 400Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile Trp 405 410 415Cys Lys Lys Pro Arg Ser Asp Lys Asn Thr Ile Ile Asp Pro Phe Asp 420 425 430Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys Met 435 440 445Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu Asn 450 455 460Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val Glu465 470 475 480Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala Leu 485 490 495Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr His Val 500 505 510Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys Tyr 515 520 525Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 530 53531780PRTBocavirus 31Met Ala Phe Asn Pro Pro Val Ile Arg Ala Phe Ser Ser Pro Ala Phe1 5 10 15Thr Tyr Val Phe Lys Phe Pro Tyr Pro Ser Trp Lys Glu Lys Glu Trp 20 25 30Leu Leu His Ala Leu Leu Ala His Gly Thr Glu Gln Ala Met Ile Gln 35 40 45Leu Arg Asn Cys Val Pro His Pro Asp Glu Asp Ile Ile Arg Asp Asp 50 55 60Leu Leu Leu Ser Leu Glu Asp Arg His Phe Gly Ala Ile Leu Cys Lys65 70 75 80Ala Val Tyr Met Ala Thr Thr Thr Phe Met Ser Gln Lys Gln Arg Asn 85 90 95Met Phe Pro Arg Cys Asp Ile Ile Val Gln Ser Glu Leu Gly Glu Thr 100 105 110Asn Leu His Cys His Ile Ile Val Gly Gly Glu Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Thr Ser Cys Pro Gln Leu Tyr Gly Leu Ile Leu Gly Glu 130 135 140Leu Ile Gln Arg Cys Lys Thr Leu Leu Ala Thr Arg Pro Phe Glu Pro145 150 155 160Glu Glu Ala Glu Ile Tyr His Ala Leu Lys Arg Ala Glu Arg Glu Ala 165 170 175Trp Gly Gly Val Thr Ser Gly Asn Leu Gln Ile Leu Gln Tyr Arg Asp 180 185 190Arg Arg Gly Asp Leu His Ala Gln Gln Val Asp Ala Leu Arg Phe Phe 195 200 205Lys Asn Tyr Leu Leu Pro Lys Asn Arg Cys Ile Thr Ser Tyr Ser Arg 210 215 220Pro Asp Val Cys Thr Ser Pro Glu Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Cys His Thr Leu Val Asn Gly Leu Pro Leu Pro Glu His Tyr 245 250 255Arg Lys His Tyr His Ala Thr Leu Asp Asn Glu Val Leu Pro Gly Pro 260 265 270Gln Thr Met Ala Phe Gly Gly Arg Gly Pro Trp Glu His Leu Pro Glu 275 280 285Val Gly Asp Gln Arg Leu Ala Ala Ser Ser Val Ser Thr Thr Tyr Lys 290 295 300Pro Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Ser305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ala Asn Cys Pro Glu 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Asn Val Cys Ser Asn Phe Thr Ala 355 360 365Leu Ser Tyr Leu Phe His Leu Tyr Pro Gly Thr Thr Leu Ser Ser Asp 370 375 380Asn Lys Ala Leu Gln Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Met385 390 395 400Val Gly His Ala Leu Cys Cys Val Leu Asn Lys Gln Phe Gly Lys Gln 405 410 415Asn Thr Val Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Met 420 425 430Ala Lys Ala Ile Val Gln Gly Ile Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Val Val 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Arg Asp 485 490 495Ser Val Leu Leu Thr Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ala Val Val Gly Gly Asn Ser Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Ile Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Thr Pro Glu Glu Ile Ala Ala Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Thr Lys 565 570 575Trp Ser Leu Asp Lys Ile Pro Asn Ser Phe Pro Leu Gly Val Leu Cys 580 585 590Pro Thr His Ser Gln Asp Phe Ile Leu His Glu Asn Gly Tyr Cys Thr 595 600 605Asp Cys Gly Gly Tyr Leu Ala His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Arg Ala Ser Asp Thr Ser Lys Glu Ala Ile Asp Ala Gly Asp Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asp Ser Glu Ser Glu Ala Ser Glu Val Gly 645 650 655Val Arg Pro Ser Lys Lys Arg Arg Ile Thr Ile Pro Ala Thr Pro Pro 660 665 670Asn Ser Pro Gly Ser Ser Val Ser Thr Ser Ala Phe Phe Asp Asn Trp 675 680 685Cys Ala Gln Pro Arg Asp Glu Asp Glu Leu Arg Glu Tyr Glu Arg Gln 690 695 700Ala Ser Arg Leu Gln Lys Lys Arg Glu Ser Arg Glu Arg Arg Glu Glu705 710 715 720Thr Pro Met Ala Thr Ser Ser Gln Glu Ser Glu Ser Glu Pro Asn Pro 725 730 735Thr Gln Trp Gly Asp Lys Leu Gly Val Ile Pro Ser Gly Thr Pro Asp 740 745 750Gln Pro Pro Ile Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp 755 760 765Glu Asp Glu Gly Glu Tyr Ile Gly Lys Glu Arg Leu 770 775 78032539PRTBocavirus 32Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Glu Pro Asn Asp1 5 10 15Gly Gln Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly Gly 20 25 30Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 35 40 45Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 50 55 60Val Lys Ile Gln Asn Asn His Gln Tyr Lys Thr Glu Asn Ile Ile Pro65 70 75 80Ser Asn Gly Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 85 90 95Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp 100 105 110Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Gly Met His 115 120 125Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 130 135 140Asp Val Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys145 150 155 160Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 165 170 175Val Met Pro Glu Leu Pro Tyr Gln Thr Trp Tyr Leu Phe Gln Tyr Gly 180 185 190Tyr Ile Pro Thr Ile His Glu Leu Ala Glu Met Glu Asp Ser Asn Ala 195 200 205Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 210 215 220Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Ala Glu Phe Asn Phe225 230 235 240Asn Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Phe Ile Pro Pro 245 250 255Gly Leu Met Phe Asn Pro Leu Val Pro Thr Arg Arg Ala Gln Tyr Ile 260 265 270Arg Arg Asn Gly Asn Thr Gln Ala Ser Thr Ser Arg Ile Gln Pro Tyr 275 280 285Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 290 295 300Arg Val Gly Pro Ala Ala Ser Asp Thr Ala Ala Trp Met Val Gly Val305 310 315 320Asp Pro Glu Gly Ala Asn Ile Asn Ser Gly Arg Ala Gly Val Ser Ser 325 330 335Gly Phe Asp Pro Pro Ala Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr 340 345 350Lys Val Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Asn Asp Gly Asn 355 360 365Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Thr Leu Tyr 370 375 380Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe385 390 395 400Pro Asn Gln Ile Trp Asp Arg Tyr Pro Val Thr Arg Glu Asn Pro Ile 405 410 415Trp Cys Lys Gln Pro Arg Ser Asp Lys His Thr Thr Ile Asp Pro Phe 420 425 430Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 435 440 445Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 450 455 460Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val465 470 475 480Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 485 490 495Leu Gly Leu Gly Ile Gly Gly Ala Asp Glu Ile Asn Pro Thr Tyr His 500 505 510Val Asp Lys Asn Gly Ala Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 515 520 525Phe Pro Val Lys Thr Asn Ile Asn Lys Val Leu 530 53533777PRTBocavirus 33Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Thr Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Asp Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asn Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610

615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Cys Asp Pro Gly Asp Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asn Ser Gln Pro Glu Thr Ser Asn Val Asp 645 650 655Asn Arg Pro Ser Lys Lys Arg Arg Val Ile Pro Glu Thr Pro Pro Asn 660 665 670Ser Pro Val Ser Arg Gln Ser Leu Ser Ser Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Glu Asp Glu Leu Arg Ile Tyr Glu Ala Gln Ala 690 695 700Ser Arg Ile Lys Glu Asn Thr Glu Ser Thr Pro Glu Arg Glu Lys Thr705 710 715 720Pro Val Gly Glu Pro Gln Glu Glu Ser Gln Ala Glu Pro Asp Pro Thr 725 730 735Ala Trp Gly Glu Lys Leu Gly Val Tyr Ser Ser Leu Gln Pro Gly Glu 740 745 750Pro Pro Ile Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu 755 760 765Asp Glu Gly Glu Asn Ile Gly Gly Glu 770 77534539PRTBocavirus 34Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Gly Ser Met Asp1 5 10 15Glu Gln Arg Gly Gly Gly Gly Gly Ala Val Gly Ser Val Gly Gly Gly 20 25 30Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 35 40 45Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 50 55 60Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro65 70 75 80Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 85 90 95Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp 100 105 110Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His 115 120 125Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 130 135 140Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys145 150 155 160Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 165 170 175Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly 180 185 190Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala 195 200 205Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 210 215 220Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe225 230 235 240Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 245 250 255Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile 260 265 270Arg Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr 275 280 285Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 290 295 300Arg Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val305 310 315 320Asn Pro Asp Gly Thr Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser 325 330 335Gly Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr 340 345 350Lys Ile Gln Trp Tyr Gln Thr Pro Glu Gly Thr Asn Ser Asp Gly Asn 355 360 365Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr 370 375 380Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe385 390 395 400Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 405 410 415Trp Cys Lys Lys Pro Arg Ser Asp Lys Ser Thr Val Ile Asp Pro Phe 420 425 430Asp Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 435 440 445Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 450 455 460Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val465 470 475 480Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 485 490 495Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Ile Asn Pro Thr Tyr His 500 505 510Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 515 520 525Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 530 53535777PRTBocavirus 35Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Asp Asp Pro Gly Asp Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asn Ser Gln Pro Glu Thr Ser Asn Val Asp 645 650 655Asn Arg Pro Ser Lys Lys Arg Arg Val Ile Pro Glu Thr Pro Pro Asn 660 665 670Ser Pro Val Ser Arg Gln Ser Leu Ser Ser Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Glu Asp Glu Leu Arg Ile Tyr Glu Ala Gln Ala 690 695 700Ser Arg Ile Lys Glu Asn Ala Glu Ser Thr Pro Glu Arg Glu Lys Thr705 710 715 720Pro Val Gly Glu Pro Gln Glu Glu Ser Gln Ser Glu Pro Asp Pro Thr 725 730 735Ala Trp Gly Glu Lys Leu Gly Val Tyr Ser Ser Leu Gln Pro Gly Glu 740 745 750Pro Pro Ile Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu 755 760 765Asp Glu Gly Glu Asn Ile Gly Gly Glu 770 77536539PRTBocavirus 36Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met Asp1 5 10 15Gly Gln Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly Gly 20 25 30Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 35 40 45Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 50 55 60Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro65 70 75 80Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 85 90 95Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp 100 105 110Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His 115 120 125Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 130 135 140Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys145 150 155 160Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 165 170 175Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly 180 185 190Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala 195 200 205Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 210 215 220Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe225 230 235 240Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 245 250 255Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile 260 265 270Arg Gln His Gly Ser Thr Ala Ser Ser Asn Thr Arg Ile Pro Pro Tyr 275 280 285Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 290 295 300Arg Val Gly Pro Ala Ala Ser Asp Ser Ala Ala Trp Met Val Val Val305 310 315 320Asn Pro Asp Gly Ala Ala Ile Asn Ser Gly Met Ala Gly Ile Gly Thr 325 330 335Gly Phe Asp Pro Pro Gly Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr 340 345 350Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly Asn 355 360 365Ile Ile Ser Asn Pro Ser Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr 370 375 380Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe385 390 395 400Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 405 410 415Trp Cys Lys Lys Pro Arg Ser Asp Lys Ser Thr Ile Ile Asp Pro Phe 420 425 430Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 435 440 445Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 450 455 460Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val465 470 475 480Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 485 490 495Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr His 500 505 510Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 515 520 525Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 530 53537777PRTBocavirus 37Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly

Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Asp Asp Pro Gly Asp Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asn Ser Gln Pro Glu Thr Ser Asn Val Asp 645 650 655Asn Arg Pro Ser Lys Lys Arg Arg Val Ile Pro Glu Thr Pro Pro Asn 660 665 670Ser Pro Val Ser Arg Gln Ser Leu Ser Ser Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Glu Asp Glu Leu Arg Ile Tyr Glu Ala Gln Ala 690 695 700Ser Arg Ile Lys Glu Asn Ala Glu Ser Thr Pro Glu Arg Glu Lys Thr705 710 715 720Pro Val Gly Glu Pro Gln Glu Glu Ser Gln Ser Glu Pro Asp Pro Thr 725 730 735Ala Trp Gly Glu Lys Leu Gly Val Tyr Ser Ser Leu Gln Pro Gly Glu 740 745 750Pro Pro Ile Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu 755 760 765Asp Glu Gly Glu Asn Ile Gly Gly Glu 770 77538539PRTBocavirus 38Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met Asp1 5 10 15Gly Gln Arg Gly Gly Gly Gly Gly Ala Ala Gly Ser Val Gly Gly Gly 20 25 30Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 35 40 45Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 50 55 60Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro65 70 75 80Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 85 90 95Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp 100 105 110Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His 115 120 125Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 130 135 140Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys145 150 155 160Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 165 170 175Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly 180 185 190Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala 195 200 205Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 210 215 220Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe225 230 235 240Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 245 250 255Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile 260 265 270Arg Gln His Gly Ser Thr Ala Ser Ser Asn Thr Arg Ile Pro Pro Tyr 275 280 285Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 290 295 300Arg Val Gly Pro Ala Ala Ser Asp Ser Ala Ala Trp Met Val Val Val305 310 315 320Asn Pro Asp Gly Ala Ala Ile Asn Ser Gly Met Ala Gly Ile Gly Thr 325 330 335Gly Phe Asp Pro Pro Gly Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr 340 345 350Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly Asn 355 360 365Ile Ile Ser Asn Pro Ser Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr 370 375 380Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe385 390 395 400Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 405 410 415Trp Cys Lys Lys Pro Arg Ser Asp Lys Ser Thr Ile Ile Asp Pro Phe 420 425 430Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 435 440 445Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 450 455 460Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val465 470 475 480Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 485 490 495Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr His 500 505 510Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 515 520 525Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 530 53539776PRTBocavirus 39Pro Pro Val Ile Arg Ala Phe Ser Ser Pro Ala Phe Thr Tyr Val Phe1 5 10 15Lys Phe Pro Tyr Arg Ser Trp Lys Glu Lys Glu Trp Leu Leu His Ala 20 25 30Leu Leu Ala His Gly Thr Glu Gln Ala Met Ile Gln Leu Arg Asn Cys 35 40 45Val Pro His Pro Asp Glu Asp Ile Ile Arg Asp Asp Leu Leu Leu Ser 50 55 60Leu Glu Asp Arg His Phe Gly Ala Ile Leu Cys Lys Ala Val Tyr Met65 70 75 80Ala Thr Thr Thr Phe Met Ser Gln Lys Gln Arg Asn Met Phe Pro Arg 85 90 95Cys Asp Ile Ile Val Gln Ser Glu Leu Gly Glu Thr Asn Leu His Cys 100 105 110His Ile Ile Val Gly Gly Glu Gly Leu Ser Lys Arg Asn Ala Lys Thr 115 120 125Ser Cys Pro Gln Leu Tyr Gly Leu Ile Leu Gly Glu Leu Ile Gln Arg 130 135 140Cys Lys Thr Leu Leu Ala Thr Arg Pro Phe Glu Pro Glu Glu Ala Glu145 150 155 160Ile Tyr His Ala Leu Lys Arg Ala Glu Arg Glu Ala Trp Gly Gly Val 165 170 175Thr Ser Gly Asn Leu Gln Ile Leu Gln Tyr Arg Asp Arg Arg Gly Asp 180 185 190Leu His Ala Gln Gln Val Asp Ala Leu Arg Phe Phe Lys Asn Tyr Leu 195 200 205Leu Pro Lys Asn Arg Cys Ile Thr Ser Tyr Ser Arg Pro Asp Val Cys 210 215 220Thr Ser Pro Glu Asn Trp Phe Val Leu Ala Glu Lys Thr Tyr Cys His225 230 235 240Thr Leu Val Asn Gly Leu Pro Leu Pro Glu His Tyr Arg Lys His Tyr 245 250 255His Ala Thr Leu Asp Asn Glu Val Leu Pro Gly Pro Gln Thr Met Ala 260 265 270Phe Gly Gly Arg Gly Pro Trp Glu His Leu Pro Glu Val Gly Asp Gln 275 280 285Arg Leu Ala Ala Ser Ser Val Ser Thr Thr Tyr Lys Pro Asn Lys Lys 290 295 300Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Ser Glu Leu Asn Leu305 310 315 320Leu Val Tyr Glu Asp Leu Val Ala Asn Cys Pro Glu Leu Leu Leu Met 325 330 335Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu Gln Val Leu Gly 340 345 350Met His His Ile Asn Val Cys Ser Asn Phe Thr Ala Leu Ser Tyr Leu 355 360 365Phe His Leu Tyr Pro Ser Thr Thr Leu Ser Ser Asp Asn Lys Ala Leu 370 375 380Gln Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Met Val Gly His Ala385 390 395 400Leu Cys Cys Val Leu Asn Lys Gln Phe Gly Lys Gln Asn Thr Val Cys 405 410 415Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Met Ala Lys Ala Ile 420 425 430Val Gln Gly Ile Arg Leu Tyr Gly Cys Val Asn His Leu Asn Lys Gly 435 440 445Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Val Val Trp Trp Glu Glu 450 455 460Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys Cys Ile Leu Gly465 470 475 480Gly Thr Glu Cys Arg Ile Asp Val Lys His Arg Asp Ser Val Leu Leu 485 490 495Thr Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp Ile Tyr Ala Val 500 505 510Val Gly Gly Asn Ser Val Ser His Val His Ala Ala Pro Leu Lys Glu 515 520 525Arg Val Ile Gln Leu Asn Phe Met Lys Gln Leu Pro Gln Thr Phe Gly 530 535 540Glu Ile Thr Pro Glu Glu Ile Ala Ala Leu Leu Gln Trp Cys Phe Asn545 550 555 560Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Thr Lys Trp Ser Leu Asp 565 570 575Lys Ile Pro Asn Ser Phe Pro Leu Gly Val Leu Cys Pro Thr His Ser 580 585 590Gln Asp Phe Ile Leu His Glu Asn Gly Tyr Cys Thr Asp Cys Gly Gly 595 600 605Tyr Leu Ala His Ser Ala Asp Asp Ser Val Tyr Thr Asp Arg Ala Ser 610 615 620Asp Thr Ser Lys Glu Ala Ile Asp Ala Gly Asp Leu Gly Asp Thr Asp625 630 635 640Gly Glu Asp Ser Glu Ser Glu Ala Ser Glu Val Gly Val Arg Pro Ser 645 650 655Lys Lys Arg Arg Ile Thr Ile Pro Ala Thr Pro Pro Asn Ser Pro Gly 660 665 670Ser Ser Val Ser Thr Ser Ala Phe Phe Asp Asn Trp Cys Ala Gln Pro 675 680 685Arg Asp Glu Asp Glu Leu Arg Glu Tyr Lys Arg Gln Ala Ser Arg Leu 690 695 700Gln Lys Lys Arg Glu Ser Arg Glu Arg Arg Glu Glu Thr Pro Met Ala705 710 715 720Thr Ser Ser Gln Glu Ser Glu Pro Glu Pro Asn Pro Thr Gln Trp Glu 725 730 735Asp Lys Leu Gly Val Ile Pro Ser Gly Thr Pro Asp Gln Pro Pro Ile 740 745 750Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu Asp Glu Gly 755 760 765Glu Tyr Ile Gly Glu Glu Arg Leu 770 77540539PRTBocavirus 40Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Glu Pro Asn Asp1 5 10 15Gly Gln Arg Gly Gly Gly Gly Gly Thr Thr Gly Ser Val Gly Gly Gly 20 25 30Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 35 40 45Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 50 55 60Val Lys Ile Gln Asn Asn His Gln Tyr Lys Thr Glu Asn Ile Ile Pro65 70 75 80Ser Asn Gly Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 85 90 95Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp 100 105 110Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Gly Met His 115 120 125Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 130 135 140Asp Val Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys145 150 155 160Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 165 170 175Val Met Pro Glu Leu Pro Tyr Gln Thr Trp Tyr Leu Phe Gln Tyr Gly 180 185 190Tyr Ile Pro Thr Ile His Glu Leu Ala Glu Met Glu Asp Ser Asn Ala 195 200 205Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 210 215 220Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Ala Glu Phe Asn Phe225 230 235 240Asn Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Phe Ile Pro Pro 245 250 255Gly Leu Met Phe Asn Pro Leu Val Pro Thr Arg Arg Ala Gln Tyr Ile 260 265 270Arg Arg Asn Gly Asn Thr Gln Ala Ser Thr Thr Arg Ile Gln Pro Tyr 275 280 285Ser Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 290 295 300Arg Val Gly Pro Ala Ala Ser Asp Thr Ala Ala Trp Met Val Gly Ile305 310 315 320Asp Leu Asp Gly Ala Asn Val Asn Ser Gly Arg Ala Gly Val Ser Thr 325 330 335Gly Phe Asp Pro Pro Ala Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr 340 345 350Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Asn Asp Gly Asn 355 360 365Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Thr Leu Tyr 370 375 380Lys Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe385 390 395 400Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 405 410 415Trp Cys Lys Gln Pro Arg Ser Asp Lys His Thr Val Ile Asp Pro Phe 420 425 430Asp Gly Ser Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 435 440 445Met Ala Lys Ile Pro Val Pro Ser Ser Asn Asn Ala Asp Ser Tyr Leu 450 455 460Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val465 470 475 480Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 485 490 495Leu Gly Leu Gly Ile Gly Gly Ala Asp Glu Ile Asn Pro Thr Tyr His 500 505 510Val Asp Lys Asn Gly Ala Tyr Ile Gln Pro Thr Ser Trp Asp Met Cys 515 520 525Phe Pro Val Lys Thr Asn Ile Asn Lys Val Leu 530 53541776PRTBocavirus 41Met Ala Phe Ser Ala Pro Val Leu Arg Ala Phe Ser Gln Pro Thr Phe1 5 10 15Thr Tyr Val Ile Lys Phe Pro Tyr Asn Asn Trp Lys Glu Asp Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Ser Leu Met Ile Gln 35 40 45Leu Lys Asn Cys Ala Pro His Pro Glu Asp Asp Pro Ile Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Leu His Tyr Gly Ala Val Phe Ala Lys65 70 75 80Ala Cys Tyr Ile Ala Thr Ser Thr Leu Met Gly Gln Lys Gln Arg Thr 85 90 95Leu Phe Pro Arg Cys Asp Ile Val Cys Gln Ser Glu Ile Gly Ser Asp 100 105 110Phe Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Arg Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala His Arg Pro Phe Glu Pro145 150 155 160Ala Glu Ala Thr Ile Tyr His Glu Leu Lys Arg Ile Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Lys Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln Pro Ile Asp Pro Leu Arg Phe Leu 195 200 205Lys His Tyr Ile Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Ser Pro Asp Asn Trp Phe Ile Leu Ala Asp Lys225 230 235 240Thr Tyr Ser His Thr Ile Ile Asn Gly Leu Pro Leu Leu Glu Arg Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Ala Met Ala Phe Gly Gly Arg Gly Ala Trp Glu Gln Leu Pro Glu 275

280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Lys His Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Phe 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Ser Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Ala Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Ile Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Phe Thr Leu His Glu Asn Gly Phe Cys Ser 595 600 605Asp Cys Gly Gly Tyr Leu Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Thr Ser Gly Asp Tyr Asp Pro Gly Asn Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asp Ser Lys Ser Glu Ala Ser Glu Val Asp 645 650 655Tyr Cys Pro Pro Lys Lys Arg Arg Val Ile Ser Ala Thr Pro Pro Asn 660 665 670Ser Pro Val Ser Gly Pro Ser Leu Ser Thr Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Asp Asp Glu Leu Arg Ile Tyr Glu Glu Gln Ala 690 695 700Ser Gln Phe Gln Lys Asn Thr Lys Ser Thr Ser Glu Arg Glu Glu Ala705 710 715 720Gln Leu Gly Glu Ser Gln Glu Pro Gln Pro Glu Pro Asp Pro Thr Ala 725 730 735Trp Gly Glu Lys Leu Gly Val Cys Ser Ser Gln Gln Pro Gly Gln Pro 740 745 750Pro Ile Val Leu Tyr Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu Asp 755 760 765Glu Gly Glu Asn Ile Gly Gly Asp 770 77542541PRTBocavirus 42Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met Asp1 5 10 15Gly Gln Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly Gly 20 25 30Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 35 40 45Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 50 55 60Val Lys Ile Gln Asn Asn His Gln Tyr Lys Thr Glu Leu Ile Ser Pro65 70 75 80Ser Thr Ser Gln Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 85 90 95Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp 100 105 110Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Gly Met His 115 120 125Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 130 135 140Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys145 150 155 160Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 165 170 175Val Met Pro Glu Leu Pro Tyr Gln Thr Trp Tyr Leu Phe Gln Tyr Gly 180 185 190Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ser Asn Ala 195 200 205Val Glu Lys Ala Ile Cys Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 210 215 220Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe225 230 235 240Asn Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 245 250 255Gly Leu Met Phe Asn Pro Leu Val Pro Thr Arg Arg Ala Gln Tyr Ile 260 265 270Arg Arg Asn Asn Asn Pro Gln Thr Ala Glu Ser Thr Ser Arg Ile Ala 275 280 285Pro Tyr Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser 290 295 300Ala Gln Arg Val Gly Pro Ala Thr Ser Asp Thr Gly Ala Trp Met Val305 310 315 320Ala Val Lys Pro Glu Asn Ala Ser Ile Asp Thr Gly Met Ser Gly Ile 325 330 335Gly Ser Gly Phe Asp Pro Pro Gln Gly Ser Leu Ala Pro Thr Asn Leu 340 345 350Glu Tyr Lys Ile Gln Trp Tyr Gln Thr Pro Gln Gly Thr Asn Asn Asn 355 360 365Gly Asn Ile Ile Ser Asn Gln Pro Leu Ser Met Leu Arg Asp Gln Ala 370 375 380Leu Phe Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp385 390 395 400Met Phe Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn 405 410 415Pro Ile Trp Cys Lys Lys Pro Arg Ser Asp Lys His Thr Thr Ile Asp 420 425 430Pro Phe Asp Gly Ser Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe 435 440 445Ile Lys Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser 450 455 460Tyr Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp465 470 475 480Glu Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His 485 490 495Thr Thr Phe Gly Leu Gly Ile Gly Gly Ala Asp Asn Leu Asn Pro Thr 500 505 510Tyr His Val Asp Lys Asn Gly Thr Tyr Ile Gln Pro Thr Thr Trp Asp 515 520 525Met Cys Phe Pro Val Lys Thr Asn Ile Asn Lys Val Leu 530 535 540435134DNABocavirus 43gacgtatgtc aaccaatcag catcgagcat atatcctata taagccaatg cacttccgca 60tctcgtcaga ctgcatccgg tctccggcga gtgaacatct ctgggaagag ctccacgcac 120gtggtgagtg acactatggc cttttctgct cctgtaatta gagctttttc tcaacctgct 180ttcacttatg ttgttaaatt tccatatgat aactggaaag aggaagagca cttactatgg 240agcttacttg ctcctgggac tgaacgtctc atgatccaac taagaaactg cgcaccacat 300cctgaagatg atcctgtcag ggaagatatt ttatgctcac tagcagacca acactatgct 360gctattttca ccaaagcttg ctacatggct gtaacttcac ttatggggca gaaacagaga 420acacactttc cacgatgcga cataatttgc caggctgaga tcggctcaga atatctacac 480tgtcacatac ttgttggagg agcaggtctg agcaagagaa atgctaaaat ttcatgtgct 540acgctcctag gccttgtgat ggctgaatta acacaacgct gcaaactact tcttgcacag 600cgtccatttg aaccagatga agctagaata tttcatctac tcagacgcgt tgaacgcgaa 660gcatggtcag ggcacactgg taactgggtt caaattcttc aatacagaga caagcgaggt 720gaccttcatg ctcaacacat tgatccttta cgctttttca aacactacct gctgccaaaa 780aatcggttga tctctccttc cagcaagcct gacgtctgca ctactccaga taactggttt 840gtcctagctg aaaaaacata cgctcacact attgttaatg ggcttccgct gctagaacat 900aacagaaagg cctatctaca agagttagaa agtgaagtca tcccagggcc ttctaccatg 960gcctttgggg gacgtggtgc gtgggaacat ctgcctgagg taggagaaca acgcctaata 1020acttctaata cttctactgc ttataaagct aacaaaaaag aaaaattaat gctaaactta 1080cttgataaat gtgatgaact taacttactt gtgtatgaag acttagttag tgcttgtcct 1140gaccttttac ttatgcttga aggtcagcca ggtggtgcac gcctaattga acaggtgctc 1200ggcatgcatc atattaaagt gtgtgctaat tacacagcgt tatctttcct atttcattta 1260catcctgatc aattattaac ttctagcaat aaagcactaa aactattgtt gattcaaggg 1320tacaacccat tgcaagtggg ccacgccatc tgttgtgtcc ttaacaaaca gatgggcaag 1380cagaacacaa tttgctttta tggccctgct tcaacaggca aaacaaatat tgcaaaggcc 1440atagttcaag gcgttcgtct gtatggctgt gttaatcatt taaacaaagg gtttgtcttt 1500aacgattgca gacaacgcct tataatctgg tgggaggagt gtttaatgca ccaagactgg 1560gtggaacctg ctaaatgcat tctaggcgga actgaatgta gaattgatgt taaacataaa 1620gacagtgttc ttcttcaaca aacaccagta attatttcca ctaaccatga catctactct 1680gtagttggtg gcaatactgt atctcatgtt catgcagcgc ccttaaaaga gcgaatcctt 1740cagctaaatt ttatgaaaca actgccacaa acatttggag aaatttctcc agttgaaatt 1800gcagaattac tgcaatggtg ctttaatgag tacgactgta ctcttactgg ctttaaacaa 1860aaatggaact tagataaagt tccaaactca tttcctcttg gggacctttg tcctacacat 1920tcacaggact acgtgcttca cgaaaacgga ttctgcactg actgcggcgg ctatattcct 1980catagtgctg acgactctgt gtacactgac gtggctagcg agacatcaat cagcagcgac 2040gacccaggta ggcattaata cattagcctt ttaatatact accttccaag tgcttatgta 2100ttaactccta caggtgactt gggggatacg gacggagaga actcccagcc ggagacatcg 2160aacgtggata atcgtccatc caagaagaga cgtgtgattc cagaaactcc accaaacagt 2220ccagtaagtc gccaaagcct ttctagcttt ttagatacgt ggcagtcaca acctagagac 2280gaagatgagc tccgaatcta tgaagcacag gcatcgcgca tcaaagagaa caccgagtcc 2340actccggaga gagagaagac accagtggga gaaccacaag aagagtcgca gtcggagccc 2400aatccgacag catggggaga aaagcttgga gtctactcct cgctacaacc aggagagccg 2460ccaatcgtct tacactgctt cgaagacctc agaccaagcg acgaagacga gggagaaaac 2520atcggggggg aatagaacca atccttatac tgtgttcagt caacacaggg ctaatcatcc 2580agatgctcct ggatggtgtg ggttttactg gcattctact aggcttgcta gagatggcac 2640taattgtatc tttaatgaaa tgaaacaaga atttcaagaa ttacaaataa atgggaaaat 2700tacttgggac aatgttagag aactattgtt tagccagaaa aaaaagctag atcaaaaata 2760cagaaacatg ctgtatcatt tcagacataa cactgattgt cctagatgtg attattggga 2820tgatgtatac cgtaaacact tagctcatgt ctcttcacag gaatcagagg aggtaacaga 2880cgaagaaatg ctttctgctg ttgaaagcat ggaaacaaat gcctccaatt aaacgccaac 2940ctggagggtg ggtgcttcct ggttataaat accttggtcc atttaatcct cttgaaaacg 3000gtgaaccagt taataaagct gatcgtgctg ctcaagctca tgataaatca tattctgaac 3060taataaaaag tggaaaaaat ccttacttat atttcaataa agctgatgaa aaattcattg 3120acgatttgaa aaacgactgg tctcttggtg gcattattgg ctcaagtttc tttaaactta 3180agcgcgccgt ggctcctgct ctaggaaata aagagcgagc tcaaaaaaga catttttact 3240ttgcaaactc aaataaaggt gctaaaaaac caaaaaataa cgagcctaaa ccaggcacat 3300caaaaatgtc tgaaaatgaa atccaagacc aacaaccatc tggctccatg gaggagcgag 3360gaggcggagg aggtgcggtc ggtagtgtgg gaggggggaa aggttctggt gtgggtatat 3420ccacargcgg ctgggttgga ggcagctact ttactgactc atatgtcata acaaagaaca 3480ctagacagtt cttagttaaa atacaaaatg accacaaata cagaacagag aatataattc 3540caagcaacgc aggaggaaaa ttccagcgat gcgtaagcac accttggtca tactttaact 3600tcaatcaata cagcagtcac ttctcaccac aagactggca gcgtttaaca aatgaatata 3660aacgctttaa gcctagaaaa atgcatgtaa aaatttacaa cttacaaata aaacaaatac 3720tctcaaatgg tgctgacact acatacaaca acgacctaac agctggtgtt cacatctttt 3780gtgatggtga acacgcatat ccaaatgcaa cacatccatg ggatgaagat gtcatgccag 3840aacttccata tgaaacatgg tatttgtttc aatatggata cattccagtt attcatgaac 3900tggctgaaat ggaagacgca aatgctgtag aaaaagctat agcactacaa atacctttct 3960tcatgcttga aaacagcgac catgaagtgt taagaacagg agaaagcaca gaattcactt 4020ttgactttga ctgtgaatgg ataaacaacg aaagagcata cattcctcct ggattaatgt 4080ttaatccaaa agttcctaca agaagagctc aatacatcag acagcacgga aacacagcat 4140ccagcaacac cagaattcaa ccatatgcaa aacctacaag ctggatgaca ggaccaggtc 4200tactcagcgc acaaagagta ggaccagctg gctcagacac tgcatcatgg atggttgttg 4260tcaatccaga cggagctgca gttaactcag gaatggcagg agttggttca ggatttgatc 4320ctccttcagg atctctaaga ccaactgact tagaatacaa aatacaatgg taccaaactc 4380ctgcaggtac caacagtgat ggaaacatca tttcaaatcc accactatcc atgctcagag 4440atcaagctct ctacagagga aatcaaacaa cctacaacct atgctcagat gtgtggatgt 4500tcccaaatca aatttgggac agatatccaa taaccagaga aaatccaatc tggtgtaaaa 4560aaccaagatc agacaaaaac acaataattg atcctttcga tggaacactt gcaatggatc 4620atccgccagg aacaatcttc ataaaaatgg caaaaattcc agttccttca aacaacaacg 4680cagactcata cctaaacatc tactgcactg gacaagtcag ctgcgaaatt gtctgggaag 4740ttgaaagata cgcaacaaag aactggagac cagaaagaag acacaccgca cttggtcttg 4800gaattggagg agaagaaaac gtaaatccaa cttatcatgt agacaaaaat ggaaaataca 4860ttcagccaac aacttgggac atgtgctatc ctatcaaaac aaacatcaat aaagtgttgt 4920aatctcttaa gcctgttcat tgcttatgct tataagttcc tctccaatgg acaagaggaa 4980agaaaagggt gactgtaatc ccgagctcat aagttcgagg ctacagtccg atggcagtgg 5040tgttgccgtc tcgaacctag ccgttacacc cttgtgcatt gtgggaggag ctgttttgct 5100tacgcaatcg cgaaatttta tatatttaat gtag 513444640PRTBocavirus 44Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys

580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Asp Asp Pro Gly Arg His625 630 635 64045777PRTBocavirus 45Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Glu Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asp Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Asp Asp Pro Gly Asp Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asn Ser Gln Pro Glu Thr Ser Asn Val Asp 645 650 655Asn Arg Pro Ser Lys Lys Arg Arg Val Ile Pro Glu Thr Pro Pro Asn 660 665 670Ser Pro Val Ser Arg Gln Ser Leu Ser Ser Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Glu Asp Glu Leu Arg Ile Tyr Glu Ala Gln Ala 690 695 700Ser Arg Ile Lys Glu Asn Thr Glu Ser Thr Pro Glu Arg Glu Lys Thr705 710 715 720Pro Val Gly Glu Pro Gln Glu Glu Ser Gln Ser Glu Pro Asn Pro Thr 725 730 735Ala Trp Gly Glu Lys Leu Gly Val Tyr Ser Ser Leu Gln Pro Gly Glu 740 745 750Pro Pro Ile Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu 755 760 765Asp Glu Gly Glu Asn Ile Gly Gly Glu 770 77546215PRTBocavirus 46Met Ser Ser Glu Ser Met Lys His Arg His Arg Ala Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Arg Arg Glu Arg Arg His Gln Trp Glu Asn His Lys 20 25 30Lys Ser Arg Ser Arg Ser Pro Ile Arg Gln His Gly Glu Lys Ser Leu 35 40 45Glu Ser Thr Pro Arg Tyr Asn Gln Glu Ser Arg Gln Ser Ser Tyr Thr 50 55 60Ala Ser Lys Thr Ser Asp Gln Ala Thr Lys Thr Arg Glu Lys Thr Ser65 70 75 80Gly Gly Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala 85 90 95Asn His Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr 100 105 110Arg Leu Ala Arg Asp Gly Thr Asn Cys Ile Phe Asn Glu Met Lys Gln 115 120 125Glu Phe Gln Glu Leu Gln Ile Asn Gly Lys Ile Thr Trp Asp Asn Val 130 135 140Arg Glu Leu Leu Phe Ser Gln Lys Lys Lys Leu Asp Gln Lys Tyr Arg145 150 155 160Asn Met Leu Tyr His Phe Arg His Asn Thr Asp Cys Pro Arg Cys Asp 165 170 175Tyr Trp Asp Asp Val Tyr Arg Lys His Leu Ala His Val Ser Ser Gln 180 185 190Glu Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser 195 200 205Met Glu Thr Asn Ala Ser Asn 210 21547667PRTBocavirusmisc_feature(170)..(170)Xaa can be any naturally occurring amino acid 47Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Glu Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Leu Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Pro Lys Asn Asn Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Gly Ser Met 130 135 140Glu Glu Arg Gly Gly Gly Gly Gly Ala Val Gly Ser Val Gly Gly Gly145 150 155 160Lys Gly Ser Gly Val Gly Ile Ser Thr Xaa Gly Trp Val Gly Gly Ser 165 170 175Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 180 185 190Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro 195 200 205Ser Asn Ala Gly Gly Lys Phe Gln Arg Cys Val Ser Thr Pro Trp Ser 210 215 220Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp225 230 235 240Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His 245 250 255Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 260 265 270Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys 275 280 285Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 290 295 300Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly305 310 315 320Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala 325 330 335Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 340 345 350Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe 355 360 365Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 370 375 380Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile385 390 395 400Arg Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr 405 410 415Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 420 425 430Arg Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val 435 440 445Asn Pro Asp Gly Ala Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser 450 455 460Gly Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr465 470 475 480Lys Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly Asn 485 490 495Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr 500 505 510Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe 515 520 525Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 530 535 540Trp Cys Lys Lys Pro Arg Ser Asp Lys Asn Thr Ile Ile Asp Pro Phe545 550 555 560Asp Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 565 570 575Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 580 585 590Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val 595 600 605Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 610 615 620Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr His625 630 635 640Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 645 650 655Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 660 66548538PRTBocavirusmisc_feature(41)..(41)Xaa can be any naturally occurring amino acid 48Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Gly Ser Met Glu1 5 10 15Glu Arg Gly Gly Gly Gly Gly Ala Val Gly Ser Val Gly Gly Gly Lys 20 25 30Gly Ser Gly Val Gly Ile Ser Thr Xaa Gly Trp Val Gly Gly Ser Tyr 35 40 45Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu Val 50 55 60Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro Ser65 70 75 80Asn Ala Gly Gly Lys Phe Gln Arg Cys Val Ser Thr Pro Trp Ser Tyr 85 90 95Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp Gln 100 105 110Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His Val 115 120 125Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala Asp 130 135 140Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys Asp145 150 155 160Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp Val 165 170 175Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly Tyr 180 185 190Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala Val 195 200 205Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn Ser 210 215 220Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe Asp225 230 235 240Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro Gly 245 250 255Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile Arg 260 265 270Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr Ala 275 280 285Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln Arg 290 295 300Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val Asn305 310 315 320Pro Asp Gly Ala Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser Gly 325 330 335Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr Lys 340 345 350Ile Gln Trp Tyr Gln Thr Pro Ala Gly Thr Asn Ser Asp Gly Asn Ile 355 360 365Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr Arg 370 375 380Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe Pro385 390 395 400Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile Trp 405 410 415Cys Lys Lys Pro Arg Ser Asp Lys Asn Thr Ile Ile Asp Pro Phe Asp 420 425 430Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys Met 435 440 445Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu Asn 450 455 460Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val Glu465 470 475 480Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala Leu 485 490 495Gly Leu Gly Ile Gly Gly Glu Glu Asn Val Asn Pro Thr Tyr His Val 500 505 510Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys Tyr 515 520 525Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 530 535495134DNABocavirus 49gacgtatgtc aaccaatcag catcgagcat atatcctata taagccgatg cacttccgca 60tctcgtcaga ctgcatccgg tctccggcga gtgaacatct ctgggaagag ctccacgcac 120gtggtgagtg acactatggc cttttctgct cctgtaatta gagctttttc tcaacctgct 180ttcacttatg ttgttaaatt tccatatgat aactggaaag aggaagaaca cttactatgg 240agcttacttg ctcctgggac tgaacgtctc atgatccaac taagaaactg cgcaccacat 300cctgaagatg atcctgtcag ggaagatatt ttatgctcac tagcagacca acactatgct 360gctattttca ccaaggcttg ctacatggct gtaacttcac tcatggggca gaaacagaga 420acacactttc cacgatgcga cataatatgc caggctgaga tcggctcaga atatctacac 480tgtcacatac ttgttggagg agcaggtcta agcaagagaa atgctaaaat ttcatgtgct 540acgctcctag gccttgtgat ggctgaatta acacaacgct gcaaactact tcttgcacag 600cgtccatttg aaccagatga agctagaata ttccatctac tcagacgcgt tgaacgcgaa 660gcatggtcag ggcacactgg taactgggtt caaattcttc aatacagaga caagcgaggt 720gaccttcatg ctcaacacat tgatccttta cgctttttca aacactacct gctgccaaaa 780aatcgattga tctctccttc cagcaagcct gacgtctgca ctactccaga taactggttt 840gtcctagctg ataaaacata cgctcacact attgttaatg ggcttccgct gctagaacat 900aacagaaagg cctatctaca agagttagag agtgaagtca tcccggggcc ttctaccatg 960gcctttgggg gacgtggtgc gtgggaacat ctgcctgagg taggagaaca acgcctaatt 1020acttctaata cttctactgc ttataaagct aacaaaaaag aaaaactaat gctaaactta 1080cttgataaat gtgatgaact taacttactt gtatatgaag

acttagttag tgcttgtcct 1140gaccttttac ttatgcttga aggtcagcca ggtggtgcac gcctaattga acaggtgcta 1200ggcatgcatc atattaaagt gtgtgctaat tatacagctc tatcattcct atttcattta 1260catcctaatc aattattaac ttctagcaat aaagcactaa aactattgtt gattcaagga 1320tacaacccat tgcaggtagg gcacgccatc tgctgtgtac ttaacaaaca gatgggcaag 1380cagaacacta tctgctttta tggtcctgct tcaacaggca aaacaaatat tgcaaaggcc 1440atagtccaag gcgttcgcct ttatggctgt gttaatcatc taaacaaagg gtttgtcttt 1500aacgattgca gacaacgcct tataatttgg tgggaggaat gtttaatgca tcaagattgg 1560gttgaacctg ctaaatgcat tttaggtgga accgaatgta gaattgatgt taaacacaaa 1620gacagtgttc ttcttcaaca aacaccagta attatttcca ctaaccatga catctactct 1680gtagttggtg gcaatactgt atctcatgtt catgcagcgc ccttaaaaga gcgaattctt 1740caactaaatt ttatgaaaca actgccacaa acatttggag agatttctcc agttgaaatt 1800gcagagttgc tgcaatggtg ctttaatgag tacgaatgta ctcttactgg ctttaaacaa 1860aaatggaact tagataaagt tccaaactca tttcctcttg gggacctttg tcctacacat 1920tcacaggact acgtgcttca cgaaaacgga ttctgcactg actgcggcgg ctatattcct 1980catagtgctg acgactctgt gtatactgac gtggctagcg agacatcaat cagcagctgc 2040gacccaggta ggcattaata cattagcctt ttaatatgct actttctaga tgcttatgta 2100ttaactccta caggtgactt gggggatacg gacggagaga actcccagcc ggagacatcg 2160aacgtggata atcgtccatc caagaagaga cgtgtgattc cagaaactcc accaaacagt 2220ccagtaagtc gccaaagcct ttctagcttt ttagatacgt ggcagtcaca acctagagac 2280gaagatgagc tccgaatcta tgaagcacag gcatcgcgca tcaaagagaa caccgagtcc 2340actccggaga gagagaagac accagtggga gaaccacaag aagagtcgca gtcggagccc 2400gatccgacag catggggaga aaagcttgga gtctactcct cgctacaacc aggagagccg 2460ccaatcgtct tacactgctt cgaagacctc agaccaagcg acgaagacga aggagaaaac 2520atcggggggg aatagaacca atccttatac tgtgttcagt caacacaggg ctaatcatcc 2580agatgctcct ggatggtgtg ggttttactg gcattctact aggcttgcta gagatggcac 2640taattgtatc tttaatgaaa tgaaacaaga atttcaagaa ttgcaaataa atggaaaaat 2700tacctgggac aatgttagag aactattgtt tagccagaaa aaaaagctag atcaaaaata 2760cagaaacatg ctgtaccatt tcagacataa tgctgattgt cctagatgtg attattggga 2820tgatgtctac cgtaaacact tagctcatgt ctcttcacag gaatcagagg aggtaacaga 2880cgaagaaatg ctttctgctg ttgaaagcat ggaaacaaat gcctccaatt aaacgccaac 2940ctggagggtg ggtgcttcct ggttataaat accttggtcc atttaatcct cttgaaaacg 3000gtaaaccagt taataaagct gatcgtgctg ctcaagctca tgataaatca tattctgaat 3060taataaagag tggaaaaaat ccttacttgt atttcaataa agctgatgag aaattcattg 3120acgatttgaa aaacgactgg tctcttggtg gcattattgg ctcaagtttc tttaaactta 3180agcgcgccgt ggctcctgct ctaggaaata aagagcgagc tcaaaaaaga catttttact 3240ttgcaaactc aaataaaggt gctaaaaaac caaaaaataa cgagcctaaa ccaggcactt 3300caaaaatgtc tgaaaatgaa atccaagacc aacaaccatc tgactcaatg gaagagcgag 3360gaggaggagg aggtgcgacc ggtagtgtgg gaggggggaa aggttctggt gtgggtatat 3420ccacaggtgg ctgggtagga ggcagctact tcactgactc atatgtcata acaaaaaaca 3480ccagacaatt tctggtaaaa atacaaaatg accacaaata cagaactgaa aatattattc 3540caagcaatgc tggaggaaaa tcacaaagat gcgtcagcac accgtggtca tatttcaact 3600tcaatcaata cagcagtcat ttttcaccac aagactggca gcgcctaaca aatgaatata 3660agcgctttaa acctagaaaa atgcatgtaa aaatttacaa tctacaaata aaacaaatac 3720tttcaaatgg tgctgacact acatacaaca acgacctaac agctggtgtt cacatctttt 3780gtgatggtga acacgcatat ccaaatgcaa cacatccatg ggatgaagac gtgatgccag 3840aacttccata tgaaacatgg tatctgtttc aatatggata cattccagtt attcatgaac 3900ttgctgaaat ggaagacgca aatgctgtag aaaaagctat agcactacaa ataccattct 3960tcatgcttga aaacagtgac catgaagttc taagaactgg agaaagcaca gaattcactt 4020ttgattttga ctgtgagtgg atcaacaacg aaagagcata cattcctcct ggattaatgt 4080ttaatccaaa agttcctacg agaagagctc aatacattag acagcacgga aacacagcat 4140caagcaacac cagaattcaa ccatatgcaa aacctacaag ctggatgaca ggaccaggtc 4200tactcagtgc acaaagagta ggaccagctg gctcagacac tgcatcatgg atggttgttg 4260ttaatccaga cggaactgcc gttaactcag gaatggcagg agttggatca ggatttgatc 4320ctccttcagg atctctaaga ccaactgact tagaatacaa aatacaatgg taccaaactc 4380ctgaaggtac caacagtgat ggaaacataa tttcaaatcc accactgtcc atgcttagag 4440atcaagctct ctacagagga aatcaaacaa cctataacct atgctcagat gtatggatgt 4500tcccaaatca aatttgggac agatatccaa taaccagaga aaacccaatt tggtgcaaaa 4560agccaagatc agataaaaac acaataattg atcctttcga tggaacactc gcaatggatc 4620atcctcctgg aacaatcttc ataaaaatgg caaaaattcc agttccttca aacaacaacg 4680cagactcata cctaaacatc tactgcacag gacaagtcag ctgcgaaatt gtctgggaag 4740ttgaaagata cgcaacaaag aactggagac cagagagaag acacaccgca cttggtcttg 4800gaatcggagg agaagaaaac ataaatccaa cttaccatgt agacaaaaat ggaaaataca 4860ttcagccaac aacatgggac atgtgctatc ctatcaaaac aaacatcaat aaagtgttgt 4920aatctcttaa gcctgttcat tgcttatgct tataagttcc tctccaatgg acaagaggaa 4980agaaaagggt gactgtaatc ccgagctcat gagttcgagg ctacagtccg atggcagtgg 5040tgttgccgtc tcgaacctag ccgttacacc cttgtgcatt gtgggaggag ctgttttgct 5100tacgcaaccg cgaaatttta tatatttaat gtag 513450640PRTBocavirus 50Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Asp Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asn Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Cys Asp Pro Gly Arg His625 630 635 64051777PRTBocavirus 51Met Ala Phe Ser Ala Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Val Lys Phe Pro Tyr Asp Asn Trp Lys Glu Glu Glu His 20 25 30Leu Leu Trp Ser Leu Leu Ala Pro Gly Thr Glu Arg Leu Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Glu Asp Asp Pro Val Arg Glu Asp 50 55 60Ile Leu Cys Ser Leu Ala Asp Gln His Tyr Ala Ala Ile Phe Thr Lys65 70 75 80Ala Cys Tyr Met Ala Val Thr Ser Leu Met Gly Gln Lys Gln Arg Thr 85 90 95His Phe Pro Arg Cys Asp Ile Ile Cys Gln Ala Glu Ile Gly Ser Glu 100 105 110Tyr Leu His Cys His Ile Leu Val Gly Gly Ala Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ile Ser Cys Ala Thr Leu Leu Gly Leu Val Met Ala Glu 130 135 140Leu Thr Gln Arg Cys Lys Leu Leu Leu Ala Gln Arg Pro Phe Glu Pro145 150 155 160Asp Glu Ala Arg Ile Phe His Leu Leu Arg Arg Val Glu Arg Glu Ala 165 170 175Trp Ser Gly His Thr Gly Asn Trp Val Gln Ile Leu Gln Tyr Arg Asp 180 185 190Lys Arg Gly Asp Leu His Ala Gln His Ile Asp Pro Leu Arg Phe Phe 195 200 205Lys His Tyr Leu Leu Pro Lys Asn Arg Leu Ile Ser Pro Ser Ser Lys 210 215 220Pro Asp Val Cys Thr Thr Pro Asp Asn Trp Phe Val Leu Ala Asp Lys225 230 235 240Thr Tyr Ala His Thr Ile Val Asn Gly Leu Pro Leu Leu Glu His Asn 245 250 255Arg Lys Ala Tyr Leu Gln Glu Leu Glu Ser Glu Val Ile Pro Gly Pro 260 265 270Ser Thr Met Ala Phe Gly Gly Arg Gly Ala Trp Glu His Leu Pro Glu 275 280 285Val Gly Glu Gln Arg Leu Ile Thr Ser Asn Thr Ser Thr Ala Tyr Lys 290 295 300Ala Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Asp305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ser Ala Cys Pro Asp 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Lys Val Cys Ala Asn Tyr Thr Ala 355 360 365Leu Ser Phe Leu Phe His Leu His Pro Asn Gln Leu Leu Thr Ser Ser 370 375 380Asn Lys Ala Leu Lys Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Gln385 390 395 400Val Gly His Ala Ile Cys Cys Val Leu Asn Lys Gln Met Gly Lys Gln 405 410 415Asn Thr Ile Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Ile 420 425 430Ala Lys Ala Ile Val Gln Gly Val Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Ile Ile 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Lys Asp 485 490 495Ser Val Leu Leu Gln Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ser Val Val Gly Gly Asn Thr Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Ile Leu Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Ser Pro Val Glu Ile Ala Glu Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Glu Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570 575Trp Asn Leu Asp Lys Val Pro Asn Ser Phe Pro Leu Gly Asp Leu Cys 580 585 590Pro Thr His Ser Gln Asp Tyr Val Leu His Glu Asn Gly Phe Cys Thr 595 600 605Asp Cys Gly Gly Tyr Ile Pro His Ser Ala Asp Asp Ser Val Tyr Thr 610 615 620Asp Val Ala Ser Glu Thr Ser Ile Ser Ser Cys Asp Pro Gly Asp Leu625 630 635 640Gly Asp Thr Asp Gly Glu Asn Ser Gln Pro Glu Thr Ser Asn Val Asp 645 650 655Asn Arg Pro Ser Lys Lys Arg Arg Val Ile Pro Glu Thr Pro Pro Asn 660 665 670Ser Pro Val Ser Arg Gln Ser Leu Ser Ser Phe Leu Asp Thr Trp Gln 675 680 685Ser Gln Pro Arg Asp Glu Asp Glu Leu Arg Ile Tyr Glu Ala Gln Ala 690 695 700Ser Arg Ile Lys Glu Asn Thr Glu Ser Thr Pro Glu Arg Glu Lys Thr705 710 715 720Pro Val Gly Glu Pro Gln Glu Glu Ser Gln Ser Glu Pro Asp Pro Thr 725 730 735Ala Trp Gly Glu Lys Leu Gly Val Tyr Ser Ser Leu Gln Pro Gly Glu 740 745 750Pro Pro Ile Val Leu His Cys Phe Glu Asp Leu Arg Pro Ser Asp Glu 755 760 765Asp Glu Gly Glu Asn Ile Gly Gly Glu 770 77552215PRTBocavirus 52Met Ser Ser Glu Ser Met Lys His Arg His Arg Ala Ser Lys Arg Thr1 5 10 15Pro Ser Pro Leu Arg Arg Glu Arg Arg His Gln Trp Glu Asn His Lys 20 25 30Lys Ser Arg Ser Arg Ser Pro Ile Arg Gln His Gly Glu Lys Ser Leu 35 40 45Glu Ser Thr Pro Arg Tyr Asn Gln Glu Ser Arg Gln Ser Ser Tyr Thr 50 55 60Ala Ser Lys Thr Ser Asp Gln Ala Thr Lys Thr Lys Glu Lys Thr Ser65 70 75 80Gly Gly Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln His Arg Ala 85 90 95Asn His Pro Asp Ala Pro Gly Trp Cys Gly Phe Tyr Trp His Ser Thr 100 105 110Arg Leu Ala Arg Asp Gly Thr Asn Cys Ile Phe Asn Glu Met Lys Gln 115 120 125Glu Phe Gln Glu Leu Gln Ile Asn Gly Lys Ile Thr Trp Asp Asn Val 130 135 140Arg Glu Leu Leu Phe Ser Gln Lys Lys Lys Leu Asp Gln Lys Tyr Arg145 150 155 160Asn Met Leu Tyr His Phe Arg His Asn Ala Asp Cys Pro Arg Cys Asp 165 170 175Tyr Trp Asp Asp Val Tyr Arg Lys His Leu Ala His Val Ser Ser Gln 180 185 190Glu Ser Glu Glu Val Thr Asp Glu Glu Met Leu Ser Ala Val Glu Ser 195 200 205Met Glu Thr Asn Ala Ser Asn 210 21553667PRTBocavirus 53Met Pro Pro Ile Lys Arg Gln Pro Gly Gly Trp Val Leu Pro Gly Tyr1 5 10 15Lys Tyr Leu Gly Pro Phe Asn Pro Leu Glu Asn Gly Lys Pro Val Asn 20 25 30Lys Ala Asp Arg Ala Ala Gln Ala His Asp Lys Ser Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asn Asp Trp Ser Leu Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Leu Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Pro Lys Asn Asn Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met 130 135 140Glu Glu Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly Gly145 150 155 160Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser

165 170 175Tyr Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu 180 185 190Val Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro 195 200 205Ser Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser 210 215 220Tyr Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp225 230 235 240Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His 245 250 255Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 260 265 270Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys 275 280 285Asp Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp 290 295 300Val Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly305 310 315 320Tyr Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala 325 330 335Val Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn 340 345 350Ser Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe 355 360 365Asp Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro 370 375 380Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile385 390 395 400Arg Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr 405 410 415Ala Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln 420 425 430Arg Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val 435 440 445Asn Pro Asp Gly Thr Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser 450 455 460Gly Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr465 470 475 480Lys Ile Gln Trp Tyr Gln Thr Pro Glu Gly Thr Asn Ser Asp Gly Asn 485 490 495Ile Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr 500 505 510Arg Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe 515 520 525Pro Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile 530 535 540Trp Cys Lys Lys Pro Arg Ser Asp Lys Asn Thr Ile Ile Asp Pro Phe545 550 555 560Asp Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys 565 570 575Met Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu 580 585 590Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val 595 600 605Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala 610 615 620Leu Gly Leu Gly Ile Gly Gly Glu Glu Asn Ile Asn Pro Thr Tyr His625 630 635 640Val Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys 645 650 655Tyr Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 660 66554538PRTBocavirus 54Met Ser Glu Asn Glu Ile Gln Asp Gln Gln Pro Ser Asp Ser Met Glu1 5 10 15Glu Arg Gly Gly Gly Gly Gly Ala Thr Gly Ser Val Gly Gly Gly Lys 20 25 30Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser Tyr 35 40 45Phe Thr Asp Ser Tyr Val Ile Thr Lys Asn Thr Arg Gln Phe Leu Val 50 55 60Lys Ile Gln Asn Asp His Lys Tyr Arg Thr Glu Asn Ile Ile Pro Ser65 70 75 80Asn Ala Gly Gly Lys Ser Gln Arg Cys Val Ser Thr Pro Trp Ser Tyr 85 90 95Phe Asn Phe Asn Gln Tyr Ser Ser His Phe Ser Pro Gln Asp Trp Gln 100 105 110Arg Leu Thr Asn Glu Tyr Lys Arg Phe Lys Pro Arg Lys Met His Val 115 120 125Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala Asp 130 135 140Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys Asp145 150 155 160Gly Glu His Ala Tyr Pro Asn Ala Thr His Pro Trp Asp Glu Asp Val 165 170 175Met Pro Glu Leu Pro Tyr Glu Thr Trp Tyr Leu Phe Gln Tyr Gly Tyr 180 185 190Ile Pro Val Ile His Glu Leu Ala Glu Met Glu Asp Ala Asn Ala Val 195 200 205Glu Lys Ala Ile Ala Leu Gln Ile Pro Phe Phe Met Leu Glu Asn Ser 210 215 220Asp His Glu Val Leu Arg Thr Gly Glu Ser Thr Glu Phe Thr Phe Asp225 230 235 240Phe Asp Cys Glu Trp Ile Asn Asn Glu Arg Ala Tyr Ile Pro Pro Gly 245 250 255Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg Ala Gln Tyr Ile Arg 260 265 270Gln His Gly Asn Thr Ala Ser Ser Asn Thr Arg Ile Gln Pro Tyr Ala 275 280 285Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu Leu Ser Ala Gln Arg 290 295 300Val Gly Pro Ala Gly Ser Asp Thr Ala Ser Trp Met Val Val Val Asn305 310 315 320Pro Asp Gly Thr Ala Val Asn Ser Gly Met Ala Gly Val Gly Ser Gly 325 330 335Phe Asp Pro Pro Ser Gly Ser Leu Arg Pro Thr Asp Leu Glu Tyr Lys 340 345 350Ile Gln Trp Tyr Gln Thr Pro Glu Gly Thr Asn Ser Asp Gly Asn Ile 355 360 365Ile Ser Asn Pro Pro Leu Ser Met Leu Arg Asp Gln Ala Leu Tyr Arg 370 375 380Gly Asn Gln Thr Thr Tyr Asn Leu Cys Ser Asp Val Trp Met Phe Pro385 390 395 400Asn Gln Ile Trp Asp Arg Tyr Pro Ile Thr Arg Glu Asn Pro Ile Trp 405 410 415Cys Lys Lys Pro Arg Ser Asp Lys Asn Thr Ile Ile Asp Pro Phe Asp 420 425 430Gly Thr Leu Ala Met Asp His Pro Pro Gly Thr Ile Phe Ile Lys Met 435 440 445Ala Lys Ile Pro Val Pro Ser Asn Asn Asn Ala Asp Ser Tyr Leu Asn 450 455 460Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile Val Trp Glu Val Glu465 470 475 480Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg Arg His Thr Ala Leu 485 490 495Gly Leu Gly Ile Gly Gly Glu Glu Asn Ile Asn Pro Thr Tyr His Val 500 505 510Asp Lys Asn Gly Lys Tyr Ile Gln Pro Thr Thr Trp Asp Met Cys Tyr 515 520 525Pro Ile Lys Thr Asn Ile Asn Lys Val Leu 530 535555217DNABocavirus 55caaggaggag tggttatatg atgtaatcca taaccactcc caggaaatga cgtatgatag 60ccaatcagaa ttgagtattg aacctatata agctgctgca cttcctgatt caatcagact 120gcatccggtc tccggcgagt gaacatctct ggaaaaagct ccacgcttgt ggtgagtcta 180ctatggcttt caatcctcct gtgattagag ctttttctca acctgctttt acttatgtct 240tcaaatttcc atatccacaa tggaaagaaa aagaatggct gcttcatgca cttttagctc 300atggaactga acaatctatg atacaattaa gaaactgcgc tcctcatccg gatgaagaca 360taatccgtga tgacttgctt atttctttag aagatcgcca ttttggggct gttctctgca 420aggctgttta catggcaaca actactctca tgtcacacaa acaaaggaat atgtttcctc 480gttgtgacat catagttcag tctgagctag gagagaaaaa cttacactgc catattatag 540ttgggggaga aggactaagc aagaggaatg ctaaatcatc ctgtgctcag ttctatggtt 600taatactagc tgaaataatt caacgctgca aatctcttct ggctacacgt ccttttgaac 660ctgaagaggc tgacatattt cacactttaa aaaaggctga gcgagaggca tggggtggag 720ttactggcgg caacatgcaa atccttcaat atagagatcg cagaggagac cttcatgcac 780aaacagtgga tcctcttcgc ttcttcaaaa actacctttt acctaaaaat agatgtattt 840catcttacag caaacctgat gtttgtactt ctcctgacaa ctggttcatt ttagctgaaa 900aaacttactc tcacactctt attaacgggc tgccgcttcc agaacattac agaaaaaact 960accacgcaac cctagataac gaagtcattc cagggcctca aacaatggcc tatggaggac 1020gtggtccgtg ggaacatctt cctgaggtag gagatcagcg cctagctgcg tcttctgtta 1080gcactactta taaacctaac aaaaaagaaa aacttatgct aaacttgcta gacaaatgta 1140aagagctaaa tctattagtt tatgaagact tagtagctaa ttgtcctgaa ctactcctta 1200tgcttgaagg tcaaccagga ggggcacgcc ttatagaaca agtcttgggc atgcaccata 1260ttaatgtttg ttctaacttt acagctctca catatctttt tcatctacat cctgttactt 1320cgcttgactc agacaataaa gctttacagc ttttgttgat tcaaggctat aatcctctag 1380ccgttggtca cgccctgtgc tgtgtcctga acaaacaatt cgggaaacaa aacactgttt 1440gcttttacgg gcctgcctca acaggtaaaa caaatatggc caaggcaatc gtccaaggga 1500ttagacttta tgggtgtgtt aatcatttga acaaaggatt tgtatttaat gactgcagac 1560aacgcttagt tgtttggtgg gaggagtgct taatgcacca ggattgggtg gaacctgcaa 1620agtgtatctt gggcgggaca gaatgcagaa ttgacgtcaa gcatagagac agtgtacttt 1680taactcaaac acctgtaatt atatccacta accacgatat ctacgcggtt gttggtggca 1740attctgtttc tcatgttcac gcggctccat taaaagaaag agtgattcag ctaaatttta 1800tgaaacaact tcctcaaaca tttggagaaa tcactgctac tgagattgca gctcttctac 1860agtggtgttt caatgagtac gactgtactc tgacaggatt taaacaaaaa tggaatttag 1920ataaaattcc aaactcattt cctcttgggg tcctttgtcc tactcattca caggacttta 1980cacttcacga aaacggatac tgcactgatt gcggtggtta ccttcctcat agtgctgaca 2040attctatgta cactgatcgc gcaagcgaaa ctagcacagg agacatcaca ccaagtaagt 2100aaatacgcat gcgcaagtaa ttcttttact ttcacttcgc tatttttacc aatttttact 2160tttaggtgac ttgggggatt cggacggaga agacaccgag cctgagacat cgcaagtgga 2220ctattgtcca cccaagaaac gtcgtctaac tgctccagca agtcctccaa actcacctgc 2280gagctctgta agtactatta ctttctttaa cacttggcac gcacagccac gtgacgaaga 2340tgagctcagg gaatatgaaa gacaagcatc gctcctacaa aagaaaaggg agtccagaaa 2400gaggggagag gaagagacac tggcagacaa ctcatcacag gagcaggagc cgcagcccga 2460tccgacacag tggggagaga ggctcgggct catatcatca ggaacaccca atcagccacc 2520tatcgtcttg cactgcttcg aagacctcag accaagtgat gaagacgagg gagagtacat 2580cggggaaaaa agacaataga acaaatccat acactgtatt cagtcaacac agagcttcca 2640atcctgaagc tccagggtgg tgtgggttct actggcactc tactcgcatt gctagagatg 2700gtactaattc aatctttaat gaaatgaaac aacagtttca acagctacaa attgataata 2760aaataggatg ggataacact agagaactat tgtttaatca aaagaaaaca ctagatcaaa 2820aatacagaaa tatgttctgg cactttagaa ataactctga ttgtgaaaga tgtaattact 2880gggatgatgt gtaccgtagg cacttagcta atgtttcctc acagacagaa gcagacgaga 2940taactgacga ggaaatgctt tctgctgctg aaagcatgga agcagatgcc tccaattaag 3000agacagccta gagggtgggt gctgcctgga tacagatatc ttgggccatt taatccactt 3060gataacggtg aacctgtaaa taacgctgat cgcgctgctc aattacatga tcacgcctac 3120tctgaactaa taaagagtgg taaaaatcca tacctgtatt tcaataaagc tgatgaaaaa 3180ttcattgatg atctaaaaga cgattggtca attggtggaa ttattggatc cagttttttt 3240aaaataaagc gcgccgtggc tcctgctctg ggaaataaag agagagccca aaaaagacac 3300ttttactttg ctaactcaaa taaaggtgca aaaaaaacaa aaaaaagtga acctaaacca 3360ggaacctcaa aaatgtctga cactgacatt caagaccaac aacctgatac tgtggacgca 3420ccacagaacg cctcaggggg aggaacagga agtattggag gaggaaaagg atctggtgtg 3480gggatttcca ctggagggtg ggtcggaggt tctcactttt cagacaaata tgtggttact 3540aaaaacacaa gacaatttat aaccacaatt cagaatggtc acctctacaa aacagaggcc 3600attgaaacaa caaaccaaag tggaaaatca cagcgctgcg tcacaactcc atggacatac 3660tttaacttta atcaatacag ctgtcacttc tcaccacaag attggcagcg ccttacaaat 3720gaatataagc gcttcagacc taaagcaatg caagtaaaga tttacaactt gcaaataaaa 3780caaatacttt caaatggtgc tgacacaaca tacaacaatg acctcacagc tggcgttcac 3840atcttttgtg atggagagca tgcttaccca aatgcatctc atccatggga tgaggacgtc 3900atgcctgatc ttccatacaa gacctggaaa ctttttcaat atggatatat tcctattgaa 3960aatgaactag cagatcttga tggaaatgca gctggaggca atgctacaga aaaagcactt 4020ctgtatcaga tgcctttttt tctacttgaa aacagtgacc accaagtact tagaactggt 4080gagagcactg aatttacttt taactttgac tgtgaatggg ttaataatga aagagcatac 4140attcctcctg gattgatgtt caatccaaaa gttccaacaa gaagagttca gtacataaga 4200caaaacggaa gcacagcagc cagcacaggc agaattcagc catactcaaa accaacaagc 4260tggatgacag gacctggcct gctcagtgca cagagagtag gaccacagtc atcagacact 4320gctccattca tggtttgcac taacccagaa ggaacacaca taaacacagg tgctgcagga 4380tttggatctg gctttgatcc tccaagcgga tgtctggcac caactaacct agaatacaaa 4440cttcagtggt accagacacc agaaggaaca ggaaataatg gaaacataat tgcaaaccca 4500tcactctcaa tgcttagaga ccaactccta tacaaaggaa accagaccac atacaatcta 4560gtgggggaca tatggatgtt tccaaatcaa gtctgggaca gatttcctat caccagagaa 4620aatccaatct ggtgcaaaaa accaagggct gacaaacaca caatcatgga tccatttgat 4680ggatccattg caatggatca tcctccaggc actattttta taaaaatggc aaaaattcca 4740gtaccaactg caacaaatgc agactcatat ctaaacatat actgtactgg acaagtcagc 4800tgtgaaattg tatgggaagt agaaagatac gcaacaaaga actggcgtcc agaaagaaga 4860catactgcac tcgggatgtc actgggagga gagagcaact acacgcctac ataccacgtg 4920gatccaacag gagcatacat ccagcccacg tcatatgatc agtgtatgcc agtaaaaaca 4980aacatcaata aagtgttgta atcttataag cctctttttt gcttctgctt acaagttcct 5040cctcaatgga caagcggaaa gtgaagggtg actgtagtcc tgagctcatg ggttcaagac 5100cacagcccga tggtagtggt gttaccgtct cgaacctagc cgacagccct tgtacattgt 5160ggggggagct gttttgtttg cttatgcaat cgcgaaactc tatatctttt aatgtgt 521756639PRTBocavirus 56Met Ala Phe Asn Pro Pro Val Ile Arg Ala Phe Ser Gln Pro Ala Phe1 5 10 15Thr Tyr Val Phe Lys Phe Pro Tyr Pro Gln Trp Lys Glu Lys Glu Trp 20 25 30Leu Leu His Ala Leu Leu Ala His Gly Thr Glu Gln Ser Met Ile Gln 35 40 45Leu Arg Asn Cys Ala Pro His Pro Asp Glu Asp Ile Ile Arg Asp Asp 50 55 60Leu Leu Ile Ser Leu Glu Asp Arg His Phe Gly Ala Val Leu Cys Lys65 70 75 80Ala Val Tyr Met Ala Thr Thr Thr Leu Met Ser His Lys Gln Arg Asn 85 90 95Met Phe Pro Arg Cys Asp Ile Ile Val Gln Ser Glu Leu Gly Glu Lys 100 105 110Asn Leu His Cys His Ile Ile Val Gly Gly Glu Gly Leu Ser Lys Arg 115 120 125Asn Ala Lys Ser Ser Cys Ala Gln Phe Tyr Gly Leu Ile Leu Ala Glu 130 135 140Ile Ile Gln Arg Cys Lys Ser Leu Leu Ala Thr Arg Pro Phe Glu Pro145 150 155 160Glu Glu Ala Asp Ile Phe His Thr Leu Lys Lys Ala Glu Arg Glu Ala 165 170 175Trp Gly Gly Val Thr Gly Gly Asn Met Gln Ile Leu Gln Tyr Arg Asp 180 185 190Arg Arg Gly Asp Leu His Ala Gln Thr Val Asp Pro Leu Arg Phe Phe 195 200 205Lys Asn Tyr Leu Leu Pro Lys Asn Arg Cys Ile Ser Ser Tyr Ser Lys 210 215 220Pro Asp Val Cys Thr Ser Pro Asp Asn Trp Phe Ile Leu Ala Glu Lys225 230 235 240Thr Tyr Ser His Thr Leu Ile Asn Gly Leu Pro Leu Pro Glu His Tyr 245 250 255Arg Lys Asn Tyr His Ala Thr Leu Asp Asn Glu Val Ile Pro Gly Pro 260 265 270Gln Thr Met Ala Tyr Gly Gly Arg Gly Pro Trp Glu His Leu Pro Glu 275 280 285Val Gly Asp Gln Arg Leu Ala Ala Ser Ser Val Ser Thr Thr Tyr Lys 290 295 300Pro Asn Lys Lys Glu Lys Leu Met Leu Asn Leu Leu Asp Lys Cys Lys305 310 315 320Glu Leu Asn Leu Leu Val Tyr Glu Asp Leu Val Ala Asn Cys Pro Glu 325 330 335Leu Leu Leu Met Leu Glu Gly Gln Pro Gly Gly Ala Arg Leu Ile Glu 340 345 350Gln Val Leu Gly Met His His Ile Asn Val Cys Ser Asn Phe Thr Ala 355 360 365Leu Thr Tyr Leu Phe His Leu His Pro Val Thr Ser Leu Asp Ser Asp 370 375 380Asn Lys Ala Leu Gln Leu Leu Leu Ile Gln Gly Tyr Asn Pro Leu Ala385 390 395 400Val Gly His Ala Leu Cys Cys Val Leu Asn Lys Gln Phe Gly Lys Gln 405 410 415Asn Thr Val Cys Phe Tyr Gly Pro Ala Ser Thr Gly Lys Thr Asn Met 420 425 430Ala Lys Ala Ile Val Gln Gly Ile Arg Leu Tyr Gly Cys Val Asn His 435 440 445Leu Asn Lys Gly Phe Val Phe Asn Asp Cys Arg Gln Arg Leu Val Val 450 455 460Trp Trp Glu Glu Cys Leu Met His Gln Asp Trp Val Glu Pro Ala Lys465 470 475 480Cys Ile Leu Gly Gly Thr Glu Cys Arg Ile Asp Val Lys His Arg Asp 485 490 495Ser Val Leu Leu Thr Gln Thr Pro Val Ile Ile Ser Thr Asn His Asp 500 505 510Ile Tyr Ala Val Val Gly Gly Asn Ser Val Ser His Val His Ala Ala 515 520 525Pro Leu Lys Glu Arg Val Ile Gln Leu Asn Phe Met Lys Gln Leu Pro 530 535 540Gln Thr Phe Gly Glu Ile Thr Ala Thr Glu Ile Ala Ala Leu Leu Gln545 550 555 560Trp Cys Phe Asn Glu Tyr Asp Cys Thr Leu Thr Gly Phe Lys Gln Lys 565 570

575Trp Asn Leu Asp Lys Ile Pro Asn Ser Phe Pro Leu Gly Val Leu Cys 580 585 590Pro Thr His Ser Gln Asp Phe Thr Leu His Glu Asn Gly Tyr Cys Thr 595 600 605Asp Cys Gly Gly Tyr Leu Pro His Ser Ala Asp Asn Ser Met Tyr Thr 610 615 620Asp Arg Ala Ser Glu Thr Ser Thr Gly Asp Ile Thr Pro Ser Lys625 630 63557219PRTBocavirus 57Met Ser Ser Gly Asn Met Lys Asp Lys His Arg Ser Tyr Lys Arg Lys1 5 10 15Gly Ser Pro Glu Arg Gly Glu Arg Lys Arg His Trp Gln Thr Thr His 20 25 30His Arg Ser Arg Ser Arg Ser Pro Ile Arg His Ser Gly Glu Arg Gly 35 40 45Ser Gly Ser Tyr His Gln Glu His Pro Ile Ser His Leu Ser Ser Cys 50 55 60Thr Ala Ser Lys Thr Ser Asp Gln Val Met Lys Thr Arg Glu Ser Thr65 70 75 80Ser Gly Lys Lys Asp Asn Arg Thr Asn Pro Tyr Thr Val Phe Ser Gln 85 90 95His Arg Ala Ser Asn Pro Glu Ala Pro Gly Trp Cys Gly Phe Tyr Trp 100 105 110His Ser Thr Arg Ile Ala Arg Asp Gly Thr Asn Ser Ile Phe Asn Glu 115 120 125Met Lys Gln Gln Phe Gln Gln Leu Gln Ile Asp Asn Lys Ile Gly Trp 130 135 140Asp Asn Thr Arg Glu Leu Leu Phe Asn Gln Lys Lys Thr Leu Asp Gln145 150 155 160Lys Tyr Arg Asn Met Phe Trp His Phe Arg Asn Asn Ser Asp Cys Glu 165 170 175Arg Cys Asn Tyr Trp Asp Asp Val Tyr Arg Arg His Leu Ala Asn Val 180 185 190Ser Ser Gln Thr Glu Ala Asp Glu Ile Thr Asp Glu Glu Met Leu Ser 195 200 205Ala Ala Glu Ser Met Glu Ala Asp Ala Ser Asn 210 21558691PRTBocavirus 58Met Ser Leu His Arg Asn Gln Arg Arg Leu Gln Thr Lys Lys Cys Phe1 5 10 15Leu Leu Leu Lys Ala Trp Ile Gln Met Pro Pro Ile Lys Arg Gln Pro 20 25 30Gly Gly Trp Val Leu Pro Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Pro 35 40 45Leu Glu Asn Gly Glu Pro Val Asn Lys Ala Asp Arg Ala Ala Gln Ala 50 55 60His Asp Lys Ser Tyr Ser Glu Ile Ile Lys Ser Gly Lys Asn Pro Tyr65 70 75 80Leu Tyr Phe Asn Lys Ala Asp Glu Lys Phe Ile Asp Asp Leu Lys Asn 85 90 95Asp Trp Ser Leu Gly Gly Ile Ile Gly Ser Ser Phe Phe Lys Leu Lys 100 105 110Arg Ala Val Ala Pro Ala Leu Gly Asn Lys Glu Arg Ala Gln Lys Arg 115 120 125His Phe Tyr Phe Ala Asn Ser Asn Lys Gly Ala Lys Lys Pro Lys Asn 130 135 140Asn Glu Pro Lys Pro Gly Thr Ser Lys Met Ser Glu Asn Glu Ile Gln145 150 155 160Asp Gln Gln Pro Ser Gly Ser Met Glu Glu Arg Gly Gly Gly Gly Gly 165 170 175Ala Val Gly Ser Val Gly Gly Gly Lys Gly Ser Ser Val Gly Ile Ser 180 185 190Thr Gly Gly Trp Val Gly Gly Ser Tyr Phe Thr Asp Ser Tyr Val Ile 195 200 205Thr Lys Asn Thr Arg Gln Phe Leu Val Lys Ile Gln Asn Asp His Lys 210 215 220Tyr Arg Thr Glu Asn Ile Ile Pro Ser Asn Ala Gly Gly Lys Ser Gln225 230 235 240Arg Cys Val Ser Thr Pro Trp Ser Tyr Phe Asn Phe Asn Gln Tyr Ser 245 250 255Ser His Phe Ser Pro Gln Asp Trp Gln His Leu Thr Asn Glu Tyr Lys 260 265 270Arg Phe Lys Pro Arg Lys Met His Val Lys Ile Tyr Asn Leu Gln Ile 275 280 285Lys Gln Ile Leu Ser Asn Gly Ala Asp Thr Thr Tyr Asn Asn Asp Leu 290 295 300Thr Ala Gly Val His Ile Phe Cys Asp Gly Glu His Ala Tyr Pro Asn305 310 315 320Ala Thr His Pro Trp Asp Glu Asp Val Met Pro Glu Leu Pro Tyr Glu 325 330 335Thr Trp Tyr Leu Phe Gln Tyr Gly Tyr Ile Pro Val Ile His Glu Leu 340 345 350Ala Glu Met Glu Asp Ala Asn Ala Val Glu Lys Ala Ile Ala Leu Gln 355 360 365Ile Pro Phe Phe Met Leu Glu Asn Ser Asp His Glu Val Leu Arg Thr 370 375 380Gly Glu Ser Thr Glu Phe Thr Phe Asp Phe Asp Cys Glu Trp Ile Asn385 390 395 400Asn Glu Arg Ala Tyr Ile Pro Pro Gly Leu Met Phe Asn Pro Lys Val 405 410 415Pro Thr Arg Arg Ala Gln Tyr Ile Arg Gln His Gly Asn Thr Ala Ser 420 425 430Ser Asn Thr Arg Ile Gln Pro Tyr Ala Lys Pro Thr Ser Trp Met Thr 435 440 445Gly Pro Gly Leu Leu Ser Ala Gln Arg Val Gly Pro Ala Gly Ser Asp 450 455 460Thr Ala Ser Trp Met Val Val Val Asn Pro Asp Gly Ala Ala Val Asn465 470 475 480Ser Gly Met Ala Gly Val Gly Ser Gly Phe Asp Pro Pro Ser Gly Ser 485 490 495Leu Arg Pro Thr Asp Leu Glu Tyr Lys Ile Gln Trp Tyr Gln Thr Pro 500 505 510Ala Gly Thr Asn Ser Asp Gly Asn Ile Ile Ser Asn Pro Pro Leu Ser 515 520 525Met Leu Arg Asp Gln Ala Leu Tyr Arg Gly Asn Gln Thr Thr Tyr Asn 530 535 540Leu Cys Ser Asp Val Trp Met Phe Pro Asn Gln Ile Trp Asp Arg Tyr545 550 555 560Pro Ile Thr Arg Glu Asn Pro Ile Trp Cys Lys Lys Pro Arg Ser Asp 565 570 575Lys Asn Thr Ile Ile Asp Pro Phe Asp Gly Thr Leu Ala Met Asp His 580 585 590Pro Pro Gly Thr Ile Phe Ile Lys Met Ala Lys Ile Pro Val Pro Ser 595 600 605Asn Asn Asn Ala Asp Ser Tyr Leu Asn Ile Tyr Cys Thr Gly Gln Val 610 615 620Ser Cys Glu Ile Val Trp Glu Val Glu Arg Tyr Ala Thr Lys Asn Trp625 630 635 640Arg Pro Glu Arg Arg His Thr Ala Leu Gly Leu Gly Ile Gly Gly Glu 645 650 655Glu Asn Val Asn Pro Thr Tyr His Val Asp Lys Asn Gly Lys Tyr Ile 660 665 670Gln Pro Thr Thr Trp Asp Met Cys Tyr Pro Ile Lys Thr Asn Ile Asn 675 680 685Lys Val Leu 69059671PRTBocavirus 59Met Pro Pro Ile Lys Arg Gln Pro Arg Gly Trp Val Leu Pro Gly Tyr1 5 10 15Arg Tyr Leu Gly Pro Phe Asn Pro Leu Asp Asn Gly Glu Pro Val Asn 20 25 30Asn Ala Asp Arg Ala Ala Gln Leu His Asp His Ala Tyr Ser Glu Leu 35 40 45Ile Lys Ser Gly Lys Asn Pro Tyr Leu Tyr Phe Asn Lys Ala Asp Glu 50 55 60Lys Phe Ile Asp Asp Leu Lys Asp Asp Trp Ser Ile Gly Gly Ile Ile65 70 75 80Gly Ser Ser Phe Phe Lys Ile Lys Arg Ala Val Ala Pro Ala Leu Gly 85 90 95Asn Lys Glu Arg Ala Gln Lys Arg His Phe Tyr Phe Ala Asn Ser Asn 100 105 110Lys Gly Ala Lys Lys Thr Lys Lys Ser Glu Pro Lys Pro Gly Thr Ser 115 120 125Lys Met Ser Asp Thr Asp Ile Gln Asp Gln Gln Pro Asp Thr Val Asp 130 135 140Ala Pro Gln Asn Ala Ser Gly Gly Gly Thr Gly Ser Ile Gly Gly Gly145 150 155 160Lys Gly Ser Gly Val Gly Ile Ser Thr Gly Gly Trp Val Gly Gly Ser 165 170 175His Phe Ser Asp Lys Tyr Val Val Thr Lys Asn Thr Arg Gln Phe Ile 180 185 190Thr Thr Ile Gln Asn Gly His Leu Tyr Lys Thr Glu Ala Ile Glu Thr 195 200 205Thr Asn Gln Ser Gly Lys Ser Gln Arg Cys Val Thr Thr Pro Trp Thr 210 215 220Tyr Phe Asn Phe Asn Gln Tyr Ser Cys His Phe Ser Pro Gln Asp Trp225 230 235 240Gln Arg Leu Thr Asn Glu Tyr Lys Arg Phe Arg Pro Lys Ala Met Gln 245 250 255Val Lys Ile Tyr Asn Leu Gln Ile Lys Gln Ile Leu Ser Asn Gly Ala 260 265 270Asp Thr Thr Tyr Asn Asn Asp Leu Thr Ala Gly Val His Ile Phe Cys 275 280 285Asp Gly Glu His Ala Tyr Pro Asn Ala Ser His Pro Trp Asp Glu Asp 290 295 300Val Met Pro Asp Leu Pro Tyr Lys Thr Trp Lys Leu Phe Gln Tyr Gly305 310 315 320Tyr Ile Pro Ile Glu Asn Glu Leu Ala Asp Leu Asp Gly Asn Ala Ala 325 330 335Gly Gly Asn Ala Thr Glu Lys Ala Leu Leu Tyr Gln Met Pro Phe Phe 340 345 350Leu Leu Glu Asn Ser Asp His Gln Val Leu Arg Thr Gly Glu Ser Thr 355 360 365Glu Phe Thr Phe Asn Phe Asp Cys Glu Trp Val Asn Asn Glu Arg Ala 370 375 380Tyr Ile Pro Pro Gly Leu Met Phe Asn Pro Lys Val Pro Thr Arg Arg385 390 395 400Val Gln Tyr Ile Arg Gln Asn Gly Ser Thr Ala Ala Ser Thr Gly Arg 405 410 415Ile Gln Pro Tyr Ser Lys Pro Thr Ser Trp Met Thr Gly Pro Gly Leu 420 425 430Leu Ser Ala Gln Arg Val Gly Pro Gln Ser Ser Asp Thr Ala Pro Phe 435 440 445Met Val Cys Thr Asn Pro Glu Gly Thr His Ile Asn Thr Gly Ala Ala 450 455 460Gly Phe Gly Ser Gly Phe Asp Pro Pro Ser Gly Cys Leu Ala Pro Thr465 470 475 480Asn Leu Glu Tyr Lys Leu Gln Trp Tyr Gln Thr Pro Glu Gly Thr Gly 485 490 495Asn Asn Gly Asn Ile Ile Ala Asn Pro Ser Leu Ser Met Leu Arg Asp 500 505 510Gln Leu Leu Tyr Lys Gly Asn Gln Thr Thr Tyr Asn Leu Val Gly Asp 515 520 525Ile Trp Met Phe Pro Asn Gln Val Trp Asp Arg Phe Pro Ile Thr Arg 530 535 540Glu Asn Pro Ile Trp Cys Lys Lys Pro Arg Ala Asp Lys His Thr Ile545 550 555 560Met Asp Pro Phe Asp Gly Ser Ile Ala Met Asp His Pro Pro Gly Thr 565 570 575Ile Phe Ile Lys Met Ala Lys Ile Pro Val Pro Thr Ala Thr Asn Ala 580 585 590Asp Ser Tyr Leu Asn Ile Tyr Cys Thr Gly Gln Val Ser Cys Glu Ile 595 600 605Val Trp Glu Val Glu Arg Tyr Ala Thr Lys Asn Trp Arg Pro Glu Arg 610 615 620Arg His Thr Ala Leu Gly Met Ser Leu Gly Gly Glu Ser Asn Tyr Thr625 630 635 640Pro Thr Tyr His Val Asp Pro Thr Gly Ala Tyr Ile Gln Pro Thr Ser 645 650 655Tyr Asp Gln Cys Met Pro Val Lys Thr Asn Ile Asn Lys Val Leu 660 665 6706030DNAArtificial SequencePrimer 60gccggagctc tgcagatatc nnnnnnnnnn 306120DNAArtificial SequencePrimer 61gccggagctc tgcagatatc 206220DNAArtificial SequencePrimer 62aacagatggg caagcagaac 206321DNAArtificial SequencePrimer 63aggacaaagg tctccaagag g 216420DNAArtificial SequencePrimer 64tgcttcaaca ggcaaaacaa 206521DNAArtificial SequencePrimer 65tccaagagga aatgagtttg g 21

Claims

1. An isolated nucleic acid molecule comprising a nucleotide sequence having at least 80% identity to SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof.

2-8. (canceled)

9. The nucleic acid of claim 1, wherein nucleotide sequence comprises an open reading frame encoding a protein selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, and conservative variants thereof.

10. (canceled)

11. A substantially purified protein encoded by a nucleotide sequence of claim 1.

12. A substantially purified protein comprising an amino acid sequence at least 80% identical to a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, and SEQ ID NO:54.

13. The protein of claim 12, comprising a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, and SEQ ID NO:54.

14. A composition comprising a protein encoded by a nucleic acid molecule of claim 1.

15. (canceled)

16. An isolated antibody that specifically binds to a protein encoded by a nucleic acid molecule of claim 1.

17. Purified serum comprising a polyclonal antibody that specifically binds to a protein encoded by a nucleic acid molecule of claim 1.

18-22. (canceled)

23. A method of detecting a Bocavirus infection in a sample comprising: a) contacting a sample suspected of containing a Bocavirus protein with an antibody that specifically binds a polypeptide encoded by SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof to form a protein/antibody complex; and b) detecting the presence of the protein/antibody complex, thereby detecting the presence of the Bocavirus protein.

24. (canceled)

25. A kit for detecting a Bocavirus in a sample, the kit comprising an antibody of claim 16.

26. (canceled)

27. A method of treating or preventing a Bocavirus infection in a subject comprising: administering to the subject an antigen encoded by a Bocavirus, the Bocavirus comprising a genome that hybridizes under highly stringent conditions to a nucleotide sequence of SEQ ID NO:1, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:43, SEQ ID NO:49, or a complement thereof; thereby treating or prevention infection in the subject.

28-33. (canceled)

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