| Entries |
| Document | Title | Date |
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| 20080286836 | METHOD AND MEANS FOR PRODUCING HIGH TITER, SAFE RECOMBINANT LENTIVIRUS VECTORS - Lentiviral vectors modified at the 5′ LTR or both the 5′ and 3′ LTR's are useful in the production of recombinant lentivirus vectors. Such vectors can be produced in the absence of a functional tat gene. Multiple transformation of the host cell with the vector carrying the transgene enhances virus production. | 11-20-2008 |
| 20080318283 | Fermentation Process for Continuous Plasmid Dna Production - A continuous process is described for the production of microbial plasmid DNA for use in biopharmaceutical and biotechnological applications. The process consists of: first growing microbial cells containing a plasmid at a reduced temperature in a continuous stage; followed by a second plasmid induction continuous culture stage with an increased temperature, with a residence time that allows accumulation of the plasmid product. A hold step at a reduced temperature after fermentation further increases the yield of plasmid product. The method enables production of a large quantity of highly purified plasmid DNA from a small bioreactor over time. | 12-25-2008 |
| 20090004703 | Method for the Production of Suitable Dna Constructs for Specific Inhibition of Gene Expression by Rna Interference - The invention relates to a method for the production of vectors which, following transfection thereof in eukaryotic cells, are suitable for targeted inhibition of the formation of defined proteins therein by RNA interference. The method for the production of such vectors does not include any PCR steps. It is a three-step procedure in a single reaction vessel and can be carried out within a few hours. Thus, a method is provided which allows very easy testing of a wide variety of siRNA sequences for their functionality within a very short time. Screening processes utilizing the rapid and uncomplicated production of vectors with the aid of said kit can be performed in a cost- and time-saving manner. Another advantage of vectors thus produced is their small size which, among other things, facilitates transfection. | 01-01-2009 |
| 20090042257 | Adenoviral fiber exchange shuttle system - The instant invention provides methods and compositions for generating recombinant adenoviral vectors. The invention also provides kits comprising for the generation of recombinant adenoviral vectors. | 02-12-2009 |
| 20090053774 | METHODS, COMPOSITIONS AND KITS FOR ONE-STEP DNA CLONING USING DNA TOPOISOMERASE - Provided herein are methods, compositions, and kits useful for molecular cloning of, for example, blunt-ended DNA molecules using DNA topoisomerase. In certain embodiments, the methods comprise combining into a mixture a first polynucleotide, wherein said first polynucleotide comprises an origin of replication, a selectable marker, two topoisomerase recognition sequences, and two nicking agent recognition sequences, wherein each of said two topoisomerase recognition sequences is within 0-50 nucleotides of at least one of the nicking agent recognition sequences and wherein each of said two nicking agent recognition sequences is nicked, with a sequence-specific topoisomerase and a second polynucleotide, wherein said second polynucleotide comprises a 5′ hydroxyl on each end of the polynucleotide; and transforming the mixture into a host organism, thereby cloning the second polynucleotide. Advantageously, in some embodiments, the methods, compositions, and kits does not require the formation or purification of a DNA-protein adduct prior to the addition of the polynucleotide to be cloned. Also provided are vector sequences to facilitate performance of the methods and methods for modifying a vector of interest to render it useful in the methods described herein. | 02-26-2009 |
| 20090123977 | System for capturing and modifying large pieces of genomic DNA and constructing organisms with synthetic chloroplasts - The functional analysis of genes frequently requires the manipulation of large genomic regions. A yeast-bacteria shuttle vector is described that can be used to clone large regions of DNA by homologous recombination. Also described is a method for isolating entire genomes, including chloroplast genomes, or large portions thereof, and manipulating the same. Also described are methods for determining minimal genomes, minimal pathway requirements, and minimal organelle genomes. | 05-14-2009 |
| 20090176283 | METHOD FOR THE OBTENTION OF CHIMERIC NUCLEOTIDE SEQUENCES AND CHIMERIC NUCLEOTIDE SEQUENCES - The present invention describes a method for producing synthetic nucleotide sequences which provides the assembly of DNA sequences, thus providing the obtention of genes, chromosomes and even whole qenomes. The method of the present invention makes use of the technique known as Polymerase Chain Reaction (PCR) but wherein no preexisting nucleic acid template is needed, being therefore an approach with minimum limitations and broad use. This method provides means for obtaining products with high industrial value, for the design and development of immunotherapeutic agents, recombinant enzymes, drugs, including the development of vaccines, gene therapy, and in applications in agriculture and environment. | 07-09-2009 |
| 20090191597 | ENHANCED PRODUCTION OF INFECTIOUS PARVOVIRUS VECTORS IN INSECT CELLS - A method of producing a packaged parvovirus vector, the method comprising: (a) providing an insect cell; (b) introducing into the insect cell one or more vectors comprising nucleotide sequences encoding: (i) a transgene flanked by TRs; and (ii) baculovirus packaging functions comprising Rep components and Cap components sufficient to result in packaging of infective parvovirus particles, wherein VP1 is supplemented relative to VP2 and VP3 sufficient to increase the production of infectious viral particles; and (c) introducing into the cell a nucleic acid encoding baculovirus helper functions for expression in the insect cell; (d) culturing the cell under conditions sufficient to produce the infectious packaged parvovirus vector. | 07-30-2009 |
| 20090253184 | COMPOSITIONS AND METHODS RELATED TO AN ADENOVIRAL TRANS-COMPLEMENTING CELL LINE - Embodiments of the invention include E1 expressing cell lines that can be used in a variety of methods for production of an E1 defective adenovirus. In certain aspects a cell of the invention can be adapted to various culture conditions, e.g., suspension culture in serum free medium. In a further aspect, the cell lines allow isolation and subculture of E1-deleted recombinant adenoviruses in an environment free of replication competent adenovirus (RCA). | 10-08-2009 |
| 20090263872 | Methods and compositions for preventing bias in amplification and sequencing reactions - The present invention is directed to compositions and methods for nucleic acid identification and detection. Compositions and methods of the present invention include template nucleic acids with stabilizing sequences. The present invention also includes concatemers formed from template nucleic acids that have stabilizing sequences, arrays of such concatemers, as well as methods for identifying and detecting sequences of such concatemers. | 10-22-2009 |
| 20090269816 | System for capturing and modifying large pieces of genomic DNA and constructing organisms with synthetic chloroplasts - The functional analysis of genes frequently requires the manipulation of large genomic regions. A yeast-bacteria shuttle vector is described that can be used to clone large regions of DNA by homologous recombination. Also described is a method for isolating entire genomes, including chloroplast genomes, or large portions thereof, and manipulating the same. Also described are methods for determining minimal genomes, minimal pathway requirements, and minimal organelle genomes. | 10-29-2009 |
| 20090275088 | PROCESSES FOR THE PREPARATION OF HIGHLY PURE PLASMID COMPOSITIONS - The present disclosure generally relates to processes for preparing highly pure plasmid compositions. The processes generally involve treating a composition comprising plasmid DNA with a polypeptide to digest colanic acid. The treated plasmid DNA is then separated from the treated composition. | 11-05-2009 |
| 20090305358 | DNA Vector Production System - The invention discloses the production of double stranded DNA (dsDNA) vectors capable of delivering nucleic acids, including cDNA, antisense, ribozyme, and small interference RNA into cells. The invention also describes nucleic acid constructs as well as methods for the production of the dsDNA vectors. | 12-10-2009 |
| 20090317875 | NUCLEIC ACID CLONING - The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families. The inventive DNA manipulation system is readily integrated with other nucleic acid manipulation systems, such as ribozyme-mediated systems, and also is susceptible to automation. | 12-24-2009 |
| 20100015669 | METHOD FOR CONSTRUCTING MICRORNA ADENOVIRUS EXPRESSION PLASMIDS OF SEVERE HEPATITIS RELATED HFGL2, HFAS AND HTNFR1 GENES AND PHARMACEUTICAL USE THEREOF - Provided is a method for constructing microRNA adenovirus expression plasmids comprising severe hepatitis related genes of hfgl2 (Human Fibrinogen-like protein 2) prothrombinase, Fas and TNFR1 (tumor necrosis factor receptor 1), and the specificity and effectivity of microRNA interference plasmids are detected at cellular level so that a combined use of three microRNA adenovirus expression plasmids for the treatment of severe hepatitis can be achieved. According to the present invention, pAd/CMV/V5-DEST vectors and pcDNA expression plasmid of hfgl2, hFas and hTNFR1 are used to construct pAd-hfgl2, pAd-hFas and pAd-hTNFR1 by means of Gateway technology. | 01-21-2010 |
| 20100047877 | Novel phospholipases and uses thereof - The invention relates to a newly identified polynucleotide sequence comprising a gene that encodes a novel phospholipase isolated from | 02-25-2010 |
| 20100055744 | DNA MINI-CIRCLES AND USES THEREOF - Methods and kits for generating circular nucleic acids in a cell-free system, and uses for the generated circular nucleic acids are provided. The methods comprise in vitro amplification of a nucleic acid template comprising a recombination site to produce tandem repeat nucleic acid sequence, and employ a recombination protein to generate the circular nucleic acids from the tandem repeat nucleic acid sequence. | 03-04-2010 |
| 20100062495 | Homologous recombination-based DNA cloning methods and compositions - Methods and compositions for cloning a donor DNA molecule into an acceptor vector at a predetermined location are described. The methods are based on homologous recombination mediated by in vitro treatment of the donor DNA and the acceptor vector with an enzyme cocktail containing an exonuclease and a single-stranded DNA binding protein. | 03-11-2010 |
| 20100136633 | COMPOSITIONS AND METHODS FOR THE ASSEMBLY OF POLYNUCLEOTIDES - The present invention provides compositions and methods for rapid assembly of one or more assembled polynucleotides from a plurality of component polynucleotides. The methods of the invention utilize circular nucleic acid vectors that comprise a DNA segment D flanked by an annealable linker sequence, annealable linker sequence pairs LA and LB, or annealable linker sequence/primer binding segment pairs LA and PB or PA and LB. Restriction endonuclease digestion of a plurality of vectors containing the DNA segments to be assembled generates a plurality of DNA fragments comprising the elements PA-D-LB, LA-D-LB, and LA-D-PB or D-LB, LA-D-LB, and LA-D. The sequences of annealable linker sequences LA and LB provide complementary termini to the DNA fragments, which are utilized in host cell mediated homologous recombination or together with promer binding segments PA and PB in a polymerase cycling assembly reaction for the ordered assembly of the various DNA segments into one or more assembled polynucleotides. | 06-03-2010 |
| 20100167357 | OBLIGATE HETERODIMER MEGANUCLEASES AND USES THEREOF - An obligate heterodimer meganuclease consisting of a first and a second monomer, deriving from two different homodimeric LAGLIDADG (SEQ ID NO: 66) endonuclease monomers (parent monomers), and having at least one pair of mutations interesting corresponding residues of said parent monomers which make an intermolecular interaction between the two monomers of each parent homodimeric LAGLIDADG (SEQ ID NO: 66) endonuclease, a vector encoding said meganuclease, a cell, an animal or a plant modified by said vector and the use of said meganuclease and derived products for molecular biology, genome engineering and genome therapy. | 07-01-2010 |
| 20100184157 | E. COLI PLASMID DNA PRODUCTION - General methods and strains of bacteria are described, that dramatically simplify and streamline plasmid DNA production. In one preferred embodiment, endolysin mediated plasmid extraction combined with flocculation mediated removal of cell debris and host nucleic acids achieves increased yield and purity with simplified downstream purification and reduced waste streams, thus reducing production costs. | 07-22-2010 |
| 20100184158 | DNA Plasmids with improved copy number - The present invention relates to the production of covalently closed circular (ccc) recombinant DNA molecules such as plasmids, cosmids, bacterial artificial chromosomes (BACs), bacteriophages, viral vectors and hybrids thereof, and more particularly to vector modifications that improve production yield of said DNA molecules in fermentation culture. | 07-22-2010 |
| 20100209974 | NON-REPLICATING PARAMYXOVIRIDAE VIRUS VECTOR - The present inventors succeeded in producing non-replicating SeV vectors whose genomic RNAs lack all genes for the NP, P, and L proteins, which are RNP-constituting proteins. The present inventors confirmed that the NP/P/L-deficient SeV vectors carrying a marker gene such as GFP provide high productivity, and high transfer and expression efficiencies of foreign genes (high MOI infection is essential for achieving high expression levels). By lacking the L gene or two or more of the NP, P, and L genes, the vectors of the present invention enable lowering the level of virus-derived proteins expressed in host cells, thereby reducing the immunogenicity upon in vivo administration. | 08-19-2010 |
| 20100304445 | NUCLEIC ACID PACKAGING SYSTEM - The present invention relates to cloning target nucleic acids using phage packaging mechanisms. Packaging initiation sites may be introduced into the target DNA. Components of a phage packaging system may be combined with the target DNA to package the DNA into phage capsids. The packaged DNA may be used to create a library of target nucleic acids, or it may be sequenced. | 12-02-2010 |
| 20110045540 | BACULOVIRUS EXPRESSION VECTOR AND METHOD THEREWITH FOR GENERATING IMMUNOGENICITY IN A HOST - A baculovirus expression vector achieves dual functions of (1) subunit vaccine by displaying the influenza surface protein for humoral immune responses; and (2) DNA vaccine by expressing influenza surface protein for long-acting cellular immune response. A method for inducing immunogenicity in a host is also disclosed. | 02-24-2011 |
| 20110165629 | Recombinase-Based Methods for Producing Expression Vectors and Compositions for Use in Practicing the Same - Methods are provided for producing a vector that includes at least one splicable intron. In the subject methods, intron containing vectors are produced from donor and acceptor vectors that each include a site specific recombinase site, where the subject donor and acceptor vectors further include splice donor and acceptor sites that, upon site specific recombination of the donor and acceptor vectors, define an intron in the product vector of the recombination step. Also provided are compositions for use in practicing the subject methods, including the donor and acceptor vectors themselves, as well as systems and kits that include the same. The subject invention finds use in a variety of different applications, including the production of expression vectors that encode C-terminal tagged fusion proteins, the production of expression vectors that encode pure protein and not a fusion thereof, and the like. | 07-07-2011 |
| 20110212493 | PERFUSION BIOREACTORS, CELL CULTURE SYSTEMS, AND METHODS FOR PRODUCTION OF CELLS AND CELL-DERIVED PRODUCTS - The present invention includes perfusion bioreactors, automated cell culture systems, and methods for production of cells and cell-derived products. | 09-01-2011 |
| 20110262974 | Novel enzymes for use in enzymatic bleaching of food products - The present invention relates to novel polypeptides according to caroase 01-05 or any functional equivalents of any of them, suitable for use in a method for preparing a food products having increased whiteness, the use of the enzyme to increase whiteness of at least part of a food product, a process for preparing a food product wherein the enzyme is used and the food product obtained. | 10-27-2011 |
| 20120083016 | METHOD FOR TRACING GRAM-NEGATIVE BACTERIA INSIDE ANIMAL MODEL USING STABLE AND BIOLUMINESCENCE-BASED EXPRESSION SYSTEM THEREFOR - A method of creating a biotechnological product and an efficient and stable bio-luminescence vector which could be used for tracking Gram-negative bacteria when distributing inside animal body are provided. Through conjugation, this auto-luminescence vector can be easily transmitted from bacteria to bacteria among Gram-negative bacteria, and may facilitate bacteria to be luminescence-labeled for subsequently analyzing the dynamic change of bio-luminescent bacteria within animal body in vivo. This system includes a lacZ promoter-driven luxABCDE, a high copy number of ColE1 replicon, and a high plasmid stability of the conjugative and broad host-ranged plasmid pSE34 from | 04-05-2012 |
| 20120129223 | DNA LOADED SUPPORTED GOLD NANOPARTICLES, PROCESS FOR THE PREPARATION AND USE THEREOF - The present invention relates to DNA loaded gold nanoparticles embedded in sharp carbonaceous carriers useful for higher DNA delivery efficiently into plants. These nanogold embedded carbon matrices are prepared by heat treatment of biogenic intracellular gold nanoparticles. The DNA delivery efficiency is tested on model plants. These materials reveal good dispersion of the transport material, producing a greater number of GUS foci per unit area. The added advantages of the composite carrier are the lower plasmid and gold requirements. Plant cell damage with the prepared carbon supported particles is very minimal and can be gauged from the increased plant regeneration and transformation efficiency compared to that of the commercial micrometer sized gold particles. This can be attributed to the sharp edges that the carbon supports possess, which lead to better piercing capabilities with minimum damage. | 05-24-2012 |
| 20120258502 | Method of producing recombinant plasmid dna using substantially solid growth medium - A method of producing recombinant plasmid DNA using substantially solid growth medium and disposable vessels in place of conventional liquid fermentation processes. The method includes inoculating a host organism containing the recombinant plasmid DNA onto the substantially solid growth medium in a disposable vessel; allowing the host organism to grow on the growth medium under conditions conducive to such growth; removing the host organism from the growth medium and lysing the host organism to access the recombinant plasmid DNA; and purifying the recombinant plasmid DNA. | 10-11-2012 |
| 20130109061 | HYPERPROLIFERATIVE RECOMBINANT CELL | 05-02-2013 |
| 20130183719 | METHOD FOR PRODUCING VIRUS VECTOR - The present invention provides a method for producing a virus vector, which comprises a step wherein cells that are capable of producing a virus vector are cultured in a culture medium that contains, as active components, a retinoic acid and a histone deacetylase inhibiting substance; and a culture medium for the production of a virus vector, which is characterized by containing, as active components, a retinoic acid and a histone deacetylase inhibiting substance. | 07-18-2013 |
| 20130203121 | METHODS FOR THE SEMI-SYNTHETIC PRODUCTION OF HIGH PURITY "MINICIRCLE" DNA VECTORS FROM PLASMIDS - The invention relates to methods and reagents for producing DNA vectors, in particular minicircle (MC) DNA vectors, in superhelical form. The invention further relates to highly pure preparations of circular DNA vectors, in particular MC DNA vectors. | 08-08-2013 |
| 20130295614 | NUCLEOTIDE SEQUENCES, METHODS, KIT AND A RECOMBINANT CELL THEREOF - The present disclosure relates to recombinant adeno-associated virus (AAV) vector serotype, wherein the capsid protein of AAV serotypes is mutated at single or multiple sites. The disclosure further relates to an improved transduction efficiency of these mutant AAV serotypes. The AAV serotypes disclosed are AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10. The instant disclosure relates to nucleotide sequences, recombinant vector, methods and kit thereof. | 11-07-2013 |
| 20140004568 | Method of Viral Production in Cells | 01-02-2014 |
| 20140051125 | Stable Production of Lentiviral Vectors - The present invention provides new stable packaging cell lines and producer cell lines as well as methods to obtain them, and a new method to produce lentiviral vectors using such cell lines. New methods and packaging cell lines of the invention are generated using a baculo-AAV hybrid system for stable expression of structural and regulatory lentiviral proteins, such system comprising a baculoviral backbone containing an integration cassette flanked by AAV ITR, in combination with a plasmid encoding rep protein. This system allows to obtain a stable integration of the structural and regulatory HIV-1 proteins gag/pol and rev. The system allows to obtain a first intermediate including only the structural and regulatory HIV proteins gag/pol and rev, to be used as starting point to obtain stable packaging cell lines as well as producer cell lines. | 02-20-2014 |
| 20140162319 | NUCLEOTIDE SEQUENCES, METHODS, KIT AND A RECOMBINANT CELL THEREOF - The present disclosure relates to recombinant adeno-associated virus (AAV) vector serotype, wherein the capsid protein of AAV serotypes is mutated at single or multiple sites. The disclosure further relates to an improved transduction efficiency of these mutant AAV serotypes. The AAV serotypes disclosed are AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10. The instant disclosure relates to nucleotide sequences, recombinant vector, methods and kit thereof. | 06-12-2014 |
| 20140170709 | VECTOR FOR GENE THERAPY - The present invention provides a retroviral vector containing a transcription unit comprising a transcription regulatory sequence and a gene encoding a polypeptide having single-stranded RNA-specific endoribonuclease activity which is placed so that its expression can be controlled by the regulatory sequence, wherein the unit is placed so that the direction of the transcription of mRNA from the unit is opposite to the direction of transcription of the RNA genome of the retroviral vector. By using the vector constructed as described above, viral supernatant showing high gene transfer efficiency can be prepared. The retroviral vector of the present invention is useful for the treatment and/or prevention of cancers and virus infections. | 06-19-2014 |
| 20140193858 | PRODUCTION OF VIRAL VECTORS - The present invention relates to methods and compositions for the production of viral vectors. In particular, the present invention provides methods and compositions for faster, higher titer and higher purity production of viral vectors (e.g. adenoviral vectors). In some embodiments, the present invention provides gutted and helper viruses with identical or similar termini. In other embodiments, the present invention provides terminal protein linked adenoviral DNA. In certain embodiments, the present invention provides template extended adenoviral DNA. | 07-10-2014 |
| 20140308711 | ENZYMATIC PHOSPHOROTHIOATION OF DNA OR RNA - This invention relates to methods, compositions, and kits for enzymatic phosphorothioation of the sugar-phosphate backbone of nucleic acids. The invention allows for phosphorothioation of pre-existing nucleic acids. | 10-16-2014 |
| 20140322760 | METHOD FOR PRODUCING VIRUS VECTOR FOR GENE TRANSFER - The present invention discloses a cell system, as a host cell to be infected with an F gene-deficient virus, which can constitutively and stably express the F protein, and a method for producing an F gene-deficient virus by utilizing the cell. A non-proliferative human parainfluenza type 2 virus vector is produced by co-culturing an F gene-deficient human parainfluenza type 2 virus with a Vero cell having the F gene of human parainfluenza type 2 virus in such a manner that the F gene is non-inducibly expressed, and isolating viral particles from a culture supernatant. | 10-30-2014 |
| 20150079636 | Circular Nucleic Acid Vectors, and Methods for Making and Using the Same - Circular nucleic acid vectors that provide for persistently high levels of protein expression are provided. The circular vectors of the subject invention are characterized by being devoid of expression-silencing bacterial sequences, where in many embodiments the subject vectors include a unidirectional site-specific recombination product hybrid sequence in addition to an expression cassette. Also provided are methods of using the subject vectors for introduction of a nucleic acid, e.g., an expression cassette, into a target cell, as well as preparations for use in practicing such methods. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications. Also provided is a highly efficient and readily scalable method for producing the vectors employed in the subject methods, as well as reagents and kits/systems for practicing the same. | 03-19-2015 |
| 20150353938 | CODON OPTIMIZED NUCLEIC ACID ENCODING A RETINITIS PIGMENTOSA GTPASE REGULATOR (RPGR) - This invention relates generally to a codon optimized nucleic acid encoding a retinitis pigmentosa GTPase regulator (RPGR) protein. The nucleic acid has enhanced stability during plasmid production relative to a wildtype cDNA encoding the RPGR protein. The invention also relates to expression cassettes, vectors, and host cells comprising the codon optimized nucleic acid. Methods for preparing a recombinant adeno-associated (rAAV) expression vector comprising the codon optimized nucleic acid sequence are also provided. The nucleic acids, expression cassettes, vectors, and host cells provided may be useful in the large scale production of rAAV expression vectors for gene therapy applications. | 12-10-2015 |
| 20150376628 | NUCLEASE-MEDIATED DNA ASSEMBLY - Methods are provided herein for assembling at least two nucleic acids using a sequence specific nuclease agent (e.g., a gRNA-Cas complex) to create end sequences having complementarity and subsequently assembling the overlapping complementary sequences. The nuclease agent (e.g., a gRNA-Cas complex) can create double strand breaks in dsDNA in order to create overlapping end sequences or can create nicks on each strand to produce complementary overhanging end sequences. Assembly using the method described herein can assemble any nucleic acids having overlapping sequences or can use a joiner oligo to assemble sequences without complementary ends. | 12-31-2015 |
| 20160032295 | METHODS, COMPOSITIONS AND KITS FOR A ONE-STEP DNA CLONING SYSTEM - Methods and kits for joining three or more polynucleotides to form a product polynucleotide are provided. Such a method includes forming a reaction mixture comprising (i) a vector fragment whose ends have four base overhangs resulting from cleavage of a vector with a first type IIs enzyme, (ii) a first insert nucleic acid with a four base overhang at one end resulting from cleavage by the first type IIs enzyme and a three base overhang at the other end resulting from cleavage by the second type IIs enzyme; and (iii) a second insert nucleic acid with a four base overhang at one end resulting from cleavage by the first type IIs enzyme and a three base overhang at the other end resulting from cleavage by the second type IIs enzyme, and (iv) a ligase. The four base overhangs of the vector ligate with the four base overhangs of the first and second inserts and the three base overhangs of the first and second inserts ligate with each other or three base overhangs of a spacer nucleic acid resulting from cleavage with the second type IIs enzyme to form a product polynucleotide in which the first and second insert nucleic acids are joined to the vector fragment. | 02-04-2016 |
| 20160122724 | Recombinant Baculovirus Expression Vector and Cell - A recombinant baculovirus expression vector or cell comprising an engineered baculovirus fp25k gene with one to three modified or mutated spots, the modified spots comprise the two 7-adenine mononucleotide repeats (MNR) and the 10 | 05-05-2016 |
| 20160130588 | METHOD FOR THE GENERATION OF POLYCYSTRONIC VECTORS - The present invention provides a method for generating polycystronic nucleic acid vectors, said method comprising the steps of a) providing a first nucleic acid vector, said first nucleic acid vector comprising: i) an origin of replication placed in front of ii) at least one first gene of interest functionally cloned within a first expression cassette, said first expression cassette comprising a promoter sequence as well as a termination sequence, said first gene of interest being located between said promoter sequence and said termination sequence, iii) a marker gene together with its termination sequence, said marker gene being situated downstream of a target sequence for a recombinase, wherein the promoter necessary for the expression of said marker gene in a host cell is absent from said first nucleic acid vector, and optionally iv) a first further marker gene which is different from the marker gene of iii), said first further marker gene being situated within said first expression cassette of ii) or within a further expression cassette, said first further marker gene allowing for the selection of cells comprising said first nucleic acid vector during the process of generating said first nucleic acid vector; b) providing a second nucleic acid vector, said second nucleic acid vector comprising i) an origin of replication placed in front of ii) at least one second gene of interest functionally cloned within a second expression cassette, said second expression cassette comprising a promoter sequence as well as a termination sequence, said second gene of interest being located between said promoter sequence and said termination sequence, iii) a promoter situated upstream of the same target sequence for a recombinase as present in front of the marker gene of iii) in the first nucleic acid vector, wherein said promoter is suitable for the expression in a host cell of said marker comprised in the first nucleic acid vector gene, and iv) optionally a second further marker gene which is different from the marker gene of iii), second further marker gene being situated within said second expression cassette of ii) or within a further expression cassette, said second further marker gene allowing for the selection of cells comprising said first nucleic acid vector during the process of generating said second nucleic acid vector; and c) contacting, under conditions suitable for a recombination to take place, a mixture of the first and the second nucleic acid vectors with a recombinase, which recombinase specifically recombines the target sequences of iii) which is situated downstream of the promoter of the second nucleic acid vector and upstream of the marker gene of the first nucleic acid vector, respectively. | 05-12-2016 |
| 20160168590 | METHODS OF IMPROVING TITER IN TRANSFECTION-BASED PRODUCTION SYSTEMS USING EUKARYOTIC CELLS | 06-16-2016 |
| 20160175455 | NANOCOMPLEX CONTAINING AMPHIPATHIC PEPTIDE USEFUL FOR EFFICIENT TRANSFECTION OF BIOMOLECULES | 06-23-2016 |
| 20160194664 | COMPOSITIONS AND METHODS FOR GENERATING ADENO-ASSOCIATED VIRAL VECTORS WITH UNDETECTABLE CAPSID GENE CONTAMINATION | 07-07-2016 |
