【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。更多还原

【Abstract】 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.更多还原