Restoration of Gene Function by Homologous Recombination: from PCR to Gene Expression in One Step
نویسندگان
چکیده
منابع مشابه
High-level transgene expression by homologous recombination-mediated gene transfer
Gene transfer and expression in eukaryotes is often limited by a number of stably maintained gene copies and by epigenetic silencing effects. Silencing may be limited by the use of epigenetic regulatory sequences such as matrix attachment regions (MAR). Here, we show that successive transfections of MAR-containing vectors allow a synergistic increase of transgene expression. This finding is par...
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PCR-based gene synthesis conventionally requires two steps: first, all overlapping oligonucleotides are assembled by self-priming; then an additional pair of primers is used to amplify the full-length gene product. Here we propose a simplified method of gene synthesis which combines these two steps into one. We have found that the efficiency of this one-step method, which we term "Simplified Ge...
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Homologous recombination (gene targeting) has many desirable features for gene therapy, because it can precisely correct mutant genes and restore their normal expression, and random nonhomologous integration of DNA is infrequent in cells in which homologous recombination has occurred. There are, however, no reports of attempts to use homologous recombination to correct mutant genes in normal he...
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ژورنال
عنوان ژورنال: Applied and Environmental Microbiology
سال: 2004
ISSN: 0099-2240,1098-5336
DOI: 10.1128/aem.70.12.7156-7160.2004