HtsA表达质粒克隆和蛋白质的纯化  

Construction of recombinant plasmid pET21d-HtsA and expression and purification of HtsA protein

作　　者：宋英莉[1] 吕春梅[1] 张晓兰[1] 边淑玲[1] 徐杰[1] 朱宛宛[1] 朱辉[1] 

机构地区：[1]哈尔滨医科大学生理学教研室,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2012年第4期320-322,328,共4页Journal of Harbin Medical University

基　　金：黑龙江省自然科学基金资助项目(LC2011C02);教育部留学回国人员科研启动基金(2011)

摘　　要：目的克隆HtsA重组表达质粒以获得HtsA表达蛋白。方法通过PCR方法扩增获得A组链球菌HtsA基因完整的序列,将此片段定向克隆到表达载体pET21d质粒中,在E.coli BL21(DE3)中表达融合蛋白,并运用离子交换层析和疏水层析分离纯化HtsA蛋白。结果经琼脂糖电泳证实成功克隆了重组表达质粒pET21d-HtsA,pET21d-HtsA融合蛋白在E.coli BL21(DE3)中得到表达,SDS-PAGE分析表明纯化的HtsA纯度较高。结论成功构建了A族链球菌HtsA基因重组表达质粒,并纯化获得了目的蛋白HtsA。Objective To clone a recombinant plasmid pET21d-HtsA for HtsA protein expres- sion and purification. Methods The target DNA fragment HtsA gene was obtained by PCR amplification and inserted into pET-21 d vector. The recombinant HtsA induced by IPTG in E. coli BL21 （ DE3 ） was purified with ion-exchange chromatography and hydrophobic chromatogra- phy and detected by SDS-PAGE. Results pET2!d-HtsA was successfully constructed by Aga- rose gel electrophoresis, the recombinant HtsA was expressed and purified. SDS-PAGE showed that the purified HtsA protein was in a high purity. Conclusion HtsA recombinant plasmid is cloned and HtsA protein is prepared successfully.

关 键 词：A组链球菌 HtsA 克隆 蛋白纯化 

分 类 号：Q78[生物学—分子生物学]