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PiggyBac Transposon Mediated Efficient eGFP Expression in Porcine Somatic Cells and Cloned Embryos

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【Author】 Luo Yi-bo1, Zhang Li1, Zhu Jiang1, Wu Mei-ling1, Huan Yan-jun1, Yin Zhi1, Mu Yan-shuang1, Xia Ping2, and Liu Zhong-hua1 1 College of Life Sciences, Northeast Agricultural University, Harbin 150030, China 2 University of California at Davis, Sacramento, California, USA

【摘要】 PiggyBac transposon has demonstrated its long-term and stable transposition on genomes of various species but lacking of the evidence on farm animal genomes. In this study, we constructed a piggyBac transposon marked with enhanced green fluorescent protein (eGFP) and showed efficient transposition in porcine somatic cells and cloned embryos. Our results demonstrated that piggyBac transposase could efficiently catalyze transposition in porcine fetal fibroblast cells, as well as in embryos. PiggyBac transposition generated 18-fold more eGFP-positive cell colonies compared to pEGFP-C1 random insertion mutagenesis, but excessive transposase might affect the transfection rate. Also piggyBac mediated 4-fold more eGFP expression than random insertion in cells and 17-fold in cloned embryos at mRNA level. When the mutagenized cells were used for somatic cell nuclear transfer (SCNT), the cleavage rate and blastocyst rate of constructed embryos harboring piggyBac transposition had no difference with random insertion group. This study provides key information on the piggyBac transposon system as a tool for creating transgenic pigs.

【基金】 Supported by the National Projects of Genetic Modified Organism Breeding Technology (2008ZX08006-002);the State Transgenic Research Programme of China (2008ZX08006-002)
  • 【分类号】S828
  • 【被引频次】3
  • 【下载频次】27
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