分子伴侣Jiv90基因的克隆及其在猪脐静脉血管内皮细胞中的表达  

Cloning of Molecular Chaperone Jiv90 Gene and Its Expression in SUVEC Line

作　　者：刘伟[1] 杨幼聪[1] 李宇立[1] 张彦明[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨陵712100

出　　处：《动物医学进展》2010年第6期26-30,共5页Progress In Veterinary Medicine

基　　金：国家转基因培育新品种专项项目(2009ZX08006006B);陕西省"13115"科技创新工程重大科技专项项目(2009ZDKG-23)

摘　　要：为研究分子伴侣Jiv90对猪瘟病毒复制的影响,克隆了分子伴侣Jiv90基因,构建了真核表达载体并在猪脐静脉血管内皮细胞中表达。根据牛的Jiv90基因序列和真核表达载体pEGFP-C1设计引物,经PCR扩增,成功克隆到猪Jiv90基因,回收后与pEGFP-C1载体连接,构建重组表达载体pEGFP-C1-Jiv90,提取质粒并采用脂质体法转染猪脐静脉血管内皮细胞。转染24 h后,荧光显微镜下可见绿色荧光蛋白表达,经G418抗性筛选得到阳性克隆。本研究成功构建了pEGFP-C1-Jiv90真核表达载体,得到稳定表达Jiv90的细胞株,为今后开展分子伴侣对猪瘟病毒复制影响的研究奠定了基础。To further explore the impact of molecular chaperone Jiv90 on classical swine fever virus replica- tion, molecular chaperone Jiv90 gene was cloned and expression plasmid was constructed and expressed in SUVEC. Primers were designed according to Jiv90 gene sequences and pEGFP-Cl. DNA fragment after PCR recovered from agrose gel was inserted into pEGFP-C1 plasmid, recombinant expression vector pEG- FP-Cl-Jiv90 was constructed. Then recombinant plasmid was transfected into SUVEC 1 line by liposome reagent, and the expression of EFGP in cells was observed by fluorescence microscopy after 24 h. The positive clones were picked out by G418 screening. The pEGFP-C1-Jivg0 was successfully constructed and Jivg0 was expressed in porcine vascular endothelial cell line,which will provide a foundation for further re- search on the impact of molecular chaperones on the virus replication.

关 键 词：猪瘟病毒 真核表达 猪脐静脉血管内皮细胞 Jiv90基因 

分 类 号：Q786[生物学—分子生物学]