摘要
目的:构建人VDAC2融合蛋白原核表达质粒并进行原核表达研究。方法:运用RT-PCR技术在培养的人HepG2.2.15细胞中钓取到目的基因VDAC2,连接至T载体上进行克隆,获得大量目的基因与原核表达载体pTrc-CKS、pMBP-P连接,构建重组融合蛋白表达质粒,转入大肠杆菌中DH5α,BL21(DE3)LySs中,IPTG诱导蛋白原核表达,表达产物经SDS-PAGE检测,分析蛋白表达情况。结果:成功构建了重组载体pTrc-CKS-VDAC2和pMBP-P-VDAC2,并在原核大肠杆菌中实现了重组融合蛋白的超量表达。结论:构建的两个人VDAC2的融合蛋白表达质粒在大肠杆菌中均得到超量表达。为进一步研究VDAC2蛋白奠定了基础。
Objective： To construct the recombinant plasmid of human VDAC2 and express the protein in E.coli. Methods： The full length of Human VDAC2 gene was cloned from human HepG2.2.15 by RT-PCR, and the VDAC2 was ligated with prokaryotic expression vector pTrc-CKS and pMBP-P respectively by recombinant DNA technique, then transform to E. coli for expression test under induction of IPTG .The expressed products were identified by SDS-PAGE. Results： Recombinant plasmid pTrc-CKS-VDAC2 and pMBP -P-VDAC2 were successfully constructed .The fusion proteins were overexpressed in E.coli. Conclusion： Human VDAC2 protein has been expressed in a great quantity in E.coli. It helps for further research the VDAC2 protein.
出处
《现代生物医学进展》
CAS
2008年第10期1805-1808,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金(30270305)
广东省自然科学基金(020733)
