中国畜牧兽医2012，Vol.39Issue(10)：145-149,5.
肝特异性表达Cre重组酶转基因猪成纤维细胞的建立
Establishment of Liver Specific Expression of Cre Recombinase Transgenic Pig Fibroblast
杨春 1朱洪伟 2刘宗岳 1王桂武 2杨福合 1邢秀梅2
作者信息
- 1. 中国农业科学院特产研究所,吉林吉林132109
- 2. 吉林省特种经济动物分子生物学重点实验室,吉林吉林132109
- 折叠
摘要
Abstract
To obtain the transgenic pig of liver specific expression Cre recombinase, we constructed porcine fibroblast line of liver specific expression Cre recombinase. The promoter of alphal-human antitrypsin and the BGHpolyA were amplified respectively from the homo-blood genome and the vector of pcDNA3. 1( + ) through PCR. Cre recombinase gene of porcine orig-ian was obtained by the method of overlap PCR. The FRT2neo cassette was digested with Xho Ⅰ and Sal Ⅰ from the vector pGC-FRT2neo, which was awarded by professor Stefano Casola of Italy. We combined the above four fragments through SOE-PCR splicing and restriction enzyme ligation, and then linked the four fragmens to the eukaryotic expression vertor pCl-neo. The recombinant was transfected into porcine fibroblast by LipofectamineTM2000. Successfully constructed the Cre recombinase expression vector and integrated to the genome of the porcine fibroblast and obtained the fibroblast of liver specific expression Cre recombinase. The obtaining of the porcine fibroblast of liver specific expression Cre recombinase should be of great value to the construction of the liver specific expression Cre recombinase transgenic pig.关键词
Cre重组酶/肝脏特异性启动子/真核表达载体/三重融合PCR/细胞转染Key words
Cre recombinase/ the promoter of liver specificity/ eukaryotic expression vector/ triplication fusion PCR/ cell transfection分类
生物科学引用本文复制引用
杨春,朱洪伟,刘宗岳,王桂武,杨福合,邢秀梅..肝特异性表达Cre重组酶转基因猪成纤维细胞的建立[J].中国畜牧兽医,2012,39(10):145-149,5.
