小鼠ZBTB9基因的克隆、亚细胞定位及原核表达  

Molecular cloning,subcellular localization and prokaryotic expression of mouse ZBTB9

作　　者：徐凤芹[1] 刘庆梅[1] 黄中开[1] 成红霞[1] 周斌[1] 车建花[1] 王尉平[1] 许维岸[1] 王明华[1] 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215123

出　　处：《扬州大学学报（农业与生命科学版）》2011年第3期15-18,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(81071957)

摘　　要：应用RT-PCR扩增技术从小鼠肺组织中成功克隆锌指蛋白ZBTB9基因;该基因含有1 380bp的开放阅读框(ORF),编码459个氨基酸。为研究其生物学功能,构建pEGFP-C1/ZBTB9融合载体转染细胞,亚细胞定位显示ZBTB9定位于细胞核且呈斑点状分布。构建原核表达载体pGEX-4T-1/ZBTB9,转入大肠杆菌BL21(DE3Lysis)中,IPTG诱导融合蛋白表达。重组蛋白经SDS-PAGE鉴定后进行纯化。结果表明:成功地纯化了ZBTB9的融合蛋白。ZBTB9基因的成功克隆及表达,为进一步深入研究ZBTB9的生物学功能奠定基础。The ZBTB9 gene was cloned from the lung cDNA of mouse by reverse transcription polymerase chain reaction,and it contains an ORF of 1 380 bp and encodes 459 amino acid protein.In order to investigate its biological function,we constructed the eukaryotic expression vector pEGFP-C1/ZBTB9.Subcellular localization analysis demonstrated that ZBTB9 was localized in the nucleus,formed dot-like structures.Constructed the prokaryotic expression vector pGEX-4T-1/ZBTB9 and expressed fusion protein in E.coli BL21（DE3 Lysis） induced by IPTG.After SDS-PAGE analysis and purification,we obtained the purified recombinant protein.In this study,the cloning and expression of ZBTB9 provided a reliable tool for the further study of its biological function.

关 键 词：锌指蛋白 ZBTB9基因 基因克隆 亚细胞定位 原核表达 

分 类 号：Q786[生物学—分子生物学]