波形蛋白基因的构建、表达及鉴定  

Construction and Expression of A Fusion Protein Vimentin

作　　者：戴丽娟[1] 李新禹[2] 陈莉[2] 唐晓波[2] 

机构地区：[1]哈尔滨医科大学附属第二医院药学部,黑龙江哈尔滨150086 [2]哈尔滨医科大学药学院生物制药学教研室,黑龙江哈尔滨150081

出　　处：《药物生物技术》2012年第1期27-30,共4页Pharmaceutical Biotechnology

基　　金：哈尔滨市科技创新人才研究专项资金项目(No.2008RFXXS018)

摘　　要：文章进行波形蛋白(VIM)基因重组质粒的构建,并进行表达与鉴定,为开辟新的肝癌诊断方法奠定基础。从人宫颈癌细胞(HELA)中提取总RNA并采用PR-PCR及PCR技术克隆VIM基因,经酶切连接等构建带有GST标签的PGEX-4T-1-VIM重组表达质粒,在大肠杆菌DH5α菌中扩增重组质粒,并在BL21菌中进行诱导表达,通过SDS-PAGE及Western blotting等进行鉴定。重组及表达蛋白Vimentin的融合蛋白,为建立一种新的肝癌早期诊断方法奠定基础。To construct a recombinant plasmids vimentin（VIM） and to express it in E.coli BL21 for establishing a method for early diagnosis of hepatocellular carcinoma（HCC）,the gene encoding VIM was amplified by RT-PCR techniques from total RNA of HELA cell,and the amplified gene was cloned into vector PGEX-4T-1 of GST tag,and transformed into DH5α.The recombinant plasmids were constructed and transformed into E.coli BL21 cells,and protein expression（including GST-Tag） was induced by IPTG and identified by SDS-PAGE and Western blotting.Fusion protein VIM with high purity could be expressed in prokaryotic system.The study has laid a foundation for opening up new ways for early diagnosis of liver cancer.

关 键 词：肝癌 VIM基因 克隆 表达 

分 类 号：Q78[生物学—分子生物学]