Patent application number | Description | Published |
20080305961 | Method of Generating Translationally Active Linear Dna Molecules and Use Thereof in Array Formats - This invention provides a method for producing expressionally and translationally active linear DNA molecules that can be used as protein expression cassettes and are useful for high throughput protein expression and analysis in living cells. The method comprises a PCR amplification using two primers complementary to the sequences flanking the DNA sequence of interest, such as a cDNA, an open reading frame or a gene, that exists in expression vectors. The resultant PCR product contains a promoter, the DNA sequence of interest, and a termination sequence. The invention further provides different uses of these translationally active DNA cassettes, such as in proteome arrays. | 12-11-2008 |
20090023592 | SYSTEM FOR IDENTIFYING AND ANALYZING EXPRESSION OF ARE-CONTAINING GENES - The present invention relates to a gene discovery system and gene expression systems specific for genes encoding ARE-containing mRNAs. In one aspect, the present invention relates to computational methods of selecting coding sequences of ARE-genes from databases using aone or more ARE search sequences. The ARE search sequences are from 10 to 80 nucleotides in length and comprise a sequence which is encompassed by one of the following two sequences: (a) WU/T(AU/TU/TU/TA)TWWW, SEQ ID NO. 1, wherein none or one of the nucleotides outside of the parenthesis is replaced by a different nucleotide, and wherein W represents A, U. or T; and (b) U/T(AU/TU/T/U/T)n, SEQ ID NO. 2, wherein n indicates that the search sequence comprises from 3 to 12 of the tetrameric sequences contained within the parenthesis. The method comprises extracting from the databases, those nucleic acids whose protein coding sequences are upstream and contiguous with a 3′untranslated region (UTR) that comprises one of the ARE search sequences. The present invention also relates to methods of selectively amplifying RNA and cDNA molecules using primers derived from and complementary to the consensus 5′ sequence motifs and primers derived from and complementary to the ARE search sequence. The present invention also relates to methods of selectively amplifying ARE genes which employ a 3′ primer which is from 15 to 50 nucleotides and length and comprises from 2 to 10 pentamers having the sequence TAAAT. The pentameric sequences in the primers are either overlapping or non-overlapping. The 3′ primers are used in the reverse transcription step of the methods, the polymerase chain reaction (PCR) amplification step of the methods, or in both the reverse transcription step and the PCR amplification step of the methods. The present invention also relates to methods of making libraries which comprise portions of the ARE genes that are selectively amplified by the present methods and to methods of making microarrays which comprise probes that hybridize under stringent conditions to portions of the protein coding sequences of the ARE genes that are selectively amplified by the present methods. The present invention also relates to libraries and the microarrays that are made by such methods. | 01-22-2009 |
20090075830 | SYSTEM FOR IDENTIFYING AND ANALYZING EXPRESSION OF ARE-CONTAINING GENES - The present invention relates to a gene discovery system and gene expression systems specific for genes encoding ARE-containing mRNAs. In one aspect, the present invention relates to computational methods of selecting coding sequences of ARE-genes from databases using a one or more ARE search sequences. The ARE search sequences are from 10 to 80 nucleotides in length and comprise a sequence which is encompassed by one of the following two sequences: (a) WU/T(AU/TU/TU/TA)TWWW, SEQ ID NO. 1, wherein none or one of the nucleotides outside of the parenthesis is replaced by a different nucleotide, and wherein W represents A, U, or T; and (b) U/T(AU/TU/T/U/T)n, SEQ ID NO. 2 wherein n indicates that the search sequence comprises from 3 to 12 of the tetrameric sequences contained within the parenthesis. The method comprises extracting from the databases, those nucleic acids whose protein coding sequences are upstream and contiguous with a 3′ untranslated region (UTR) that comprises one of the ARE search sequences. The present invention also relates to methods of selectively amplifying RNA and cDNA molecules using primers derived from and complementary to the consensus 5′ sequence motifs and primers derived from and complementary to the ARE search sequence. The present invention also relates to methods of selectively amplifying ARE genes which employ a 3′ primer which is from 15 to 50 nucleotides and length and comprises from 2 to 10 pentamers having the sequence TAAAT. The pentameric sequences in the primers are either overlapping or non-overlapping. The 3′ primers are used in the reverse transcription step of the methods, the polymerase chain reaction (PCR) amplification step of the methods, or in both the reverse transcription step and the PCR amplification step of the methods. The present invention also relates to methods of making libraries which comprise portions of the ARE genes that are selectively amplified by the present methods and to methods of making microarrays which comprise probes that hybridize under stringent conditions to portions of the protein coding sequences of the ARE genes that are selectively amplified by the present methods. The present invention also relates to libraries and the microarrays that are made by such methods. | 03-19-2009 |
20090181427 | HYBRID 3' UNTRANSLATED REGIONS SUITABLE FOR EFFICIENT PROTEIN EXPRESSION IN MAMMALIAN CELLS - The present invention describes the use of hybrid short 3′ untranslated (3′UTR) regions which are composed of two regions, one region from an 3′ untranslated region of a stable eukaryotic mRNA, and another region from the downstream end of an 3′ untranslated region of another eukaryotic mRNA that contains a polyadenylation (polyA) signal. The use of such hybrid regions allows for an efficient protein expression when used in conjunction with circular or linear expression DNA molecules. The present invention provides a recombinant DNA that is composed of a promoter, a protein coding region, and the hybrid 3′UTR in a continuous and directional orientation. The efficient expression systems of the invention are suitable for the economical and efficient production of therapeutic proteins and for use with both transient and stable expression systems. | 07-16-2009 |
20110166042 | Methods for Producing Inducible and/or Repressible Expression Active Linear RNA Interference Cassettes and Inducible and/or Repressible Expression Active Linear Gene Cassettes and Their Uses - The present invention relates to a (first) method for producing inducible and/or repressible expression active linear RNA interference constructs comprising a PCR amplification of a source polynucleotide comprising the inhibitory RNA coding sequence of interest or comprising a PCR amplification of a DNA source comprising a promoter using a reverse primer comprising the inhibitory RNA coding sequence of interest. The present invention furthermore relates to a (second) method for producing inducible and/or repressible expression active linear gene constructs comprising a PCR amplification of a source expression polynucleotide comprising a promoter sequence and the DNA sequence of interest or comprising a PCR amplification using the DNA sequence as a template. The present invention furthermore relates to libraries, arrays, cells and cell lines and kits utilizing the inducible and/or repressible expression active linear RNA interference constructs or the inducible and/or repressible expression active linear gene constructs. | 07-07-2011 |
20110312543 | Expression Vectors Based on Modified Ribosomal Protein Promoters and Uses Thereof in Post-Transcriptional Assessment - The present invention relates to expression vector comprising (a) a promoter region comprising a non-inducible constitutively active ribosomal protein gene promoter, (b) an operably linked reporter or gene sequence, and (c) a 3′ untranslated region (3′ UTR), which are suitable means for an selective assessment of post-transcriptional regulation, post-transcriptional control elements and factors as well as for identifying compounds that effect post-transcription. The present invention furthermore relates to arrays, expression vector libraries and cell lines containing the expression vector(s). The present invention furthermore relates to a method and kit for identifying compounds that affect post-transcriptional regulation of reporter(s) or gene(s), that utilize the expression vector(s). | 12-22-2011 |
20130203104 | FLUORESCENT PROTEINS WITH INCREASED ACTIVITY IN CELLS - The present invention relates to fluorescent proteins, in particular green fluorescent proteins (GFPs), with increased activity in cells, and thus increased signal strength. A further aspect of the present invention relates to the use of peptides for increasing the expression and/or stability of a protein in a cell. | 08-08-2013 |
20140295544 | FLUORESCENT PROTEINS WITH INCREASED ACTIVITY IN CELLS - The present invention relates to fluorescent proteins, in particular green fluorescent proteins (GFPs), with increased activity in cells, and thus increased signal strength. A further aspect of the present invention relates to the use of peptides for increasing the expression and/or stability of a protein in a cell. | 10-02-2014 |