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- W2353121278 abstract "Efficient and targeted genome modification is now the hot topic in gene engineering animal.Streptomyces ΦC31 integrase is capable of integrating attB-containing plasmid into pseudo attP site in mammalian genomes,resulting in the normal structure and high expression of transgene.This study is to discover the molecular basis of ΦC31 integrase-mediated site-specific transgene integration in pig genome.A reporter plasmid pEGFP-N1-attB was constructed and co-transfected with ΦC31 integraseexpressing plasmid pCMV-INT into pig kidney PK15 cells.Cell clones were generated under G418 selection.Quantitative real-time PCR was used to identify cell clones with single-copy transgene integration.One pig pseudo attP site,pig-attP-1 was isolated by TAIL-PCR.It is located in an intergenic region in pig chromosome 1,spanning from 114220087-114220126.Sanger sequencing showed that pEGFP-N1-attP was broken at attB site and joined with pig-attP-1 site.Extracellular EGFP expression was measured by a fluorescence detector,where its expression level was 50 fold higher than that in background fluorescence(13500AU vs.280AU),implying that pig-attP-1 site is a safe harbor in favor of transgene expression.This study paves a new way for site-specific transgene integration in pig genome.It also provides a new strategy for creating genetically modified pig and animal bioreactor." @default.
- W2353121278 created "2016-06-24" @default.
- W2353121278 creator A5080677639 @default.
- W2353121278 date "2014-01-01" @default.
- W2353121278 modified "2023-09-24" @default.
- W2353121278 title "Site-specific Modification of Pig Genome Mediated by ΦC31 Integrase" @default.
- W2353121278 hasPublicationYear "2014" @default.
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