摘要
目的:构建hLMO4的原核表达载体并诱导、纯化和鉴定其表达。方法:hLMO4全长编码基因经BamHⅠ和XhoⅠ双酶切后,克隆至GST融合表达载体pGEX-5X-1,异丙基β-D硫代半乳糖苷(IPTG)在BL21大肠杆菌中诱导GST-hLMO4融合蛋白表达,利用Glutathione Sepharose 4B纯化诱导的融合蛋白,并经Western blot鉴定结果。结果:hLMO4编码序列克隆至pGEX-5X-1载体中,双酶切鉴定片段大小为500 bp,在E.coli BL21中IPTG诱导融合蛋白的表达,分子质量约为49 000 Da,成功纯化出GST及GST-hLMO4蛋白,Western blot检测到蛋白表达。结论:构建了hLMO4基因原核表达载体,鉴定了GST-hLMO4融合蛋白表达。
Objective：To construct prokaryotic expression vector of human LMO4 gene and induce,purify and identify its recombinant protein expression.Methods：The hLMO4 coding sequence was digested with BamHⅠ and XhoⅠ enzymes,and cloned into pGEX-5X-1.The expression of GST-hLMO4 fusion protein was induced by IPTG and identified by Western blot.Results：The coding sequence of hLMO4 gene was cloned into the pGEX-5X-1 plasmid which was transformed into E.coli BL21.The length of fragment was 500 bp.The expression of GST-hLMO4 fusion protein was induced by IPTG,and the molecular weight of protein was 49 000 Da.Conclusion：The recombinant prokaryotic plasmid was successfully constructed into pGEX-5X-1.The expression of GST-LMO4 fusion protein was induced by IPTG and identified.
出处
《东南大学学报（医学版）》
CAS
2012年第2期139-143,共5页
Journal of Southeast University(Medical Science Edition)
基金
国家自然科学基金资助项目(31171360
30800415)
博士点基金资助项目(20102104110016)
关键词
LMO4
蛋白纯化
融合蛋白
LMO 4
protein purification
fusion protein
