肝特异性表达Cre重组酶转基因猪成纤维细胞的建立

Abstract

Abstract: To obtain the transgenic swine of liver specific expression Cre recombinase, we constructed porcine fibroblast line of liver specific expression Cre recombinase. The promoter of alpha1-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 origian was obtained by the method of overlap PCR. The FRT2neo cassette was digested with XhoⅠ and SalⅠ 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 pC1-neo. The recombinant was transfected into porcine fibroblast by Lipofectamine^TM2000. 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 swine.

Key words: Cre recombinase; the promoter of liver specificity; eukaryotic expression vector; triplication fusion PCR; cell transfection

CLC Number:

Q813

YANG Chun, ZHU Hong-wei, LIU Zong-yue, WANG Gui-wu, YANG Fu-he, XING Xiu-mei. Establishment of Liver Specific Expression of Cre Recombinase Transgenic Pig Fibroblast[J]. , 2012, 39(10): 145-149.

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Establishment of Liver Specific Expression of Cre Recombinase Transgenic Pig Fibroblast

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