摘要
[目的]构建河南华溪蟹(Sinopotamon henanense)金属硫蛋白(MT)的原核表达载体。[方法]以大肠杆菌DH-5α中的pGEM-MT质粒为模板,PCR扩增目的片段并连接至pGEM-T载体。然后,亚克隆至pQE31表达载体,转化大肠杆菌DH-5α感受态细胞。[结果]PCR扩增目的片段产物大小约为220 bp,与预期大小一致。重组质粒pQE31-MT双酶切鉴定结果也与预期相符。[结论]成功构建了MT原核表达载体,为今后表达并纯化MT提供了材料。
[Objective] To construct the prokaryotic expression vector for metallothionein（MT） of Sinopotamon henanense.[Method] The plasmid pGEM-MT from E.coli DH-5α was used as the template.The target fragment of MT gene was amplified using PCR,and then bound to the vector pGEM-MT for amplification.The amplified fragment of the MT gene was subcloned into the prokaryotic expression vector pQE31 and then transferred to E.coli DH-5α.[Result] DNA fragment in size of 220 bp were amplified.The size of the PCR products met our expectation and the results of recombinant plasmid pQE31-MT double-enzyme cleavage was in line with the expectation.[Conclusion] A prokaryotic expression vector for MT has been successfully constructed,provided materials for the expression and the purification of the protein.
出处
《安徽农业科学》
CAS
北大核心
2011年第26期16132-16133,共2页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(30870267与30970361)
山西省自然科学基金项目(2008011069)
山西省回国留学人员科研项目
关键词
金属硫蛋白
基因重组
表达载体pQE31
原核表达
Metallothionein
Gene recombination
Expression vector pQE31
Prokaryotic expression
