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HtsA表达质粒克隆和蛋白质的纯化 被引量：1

Construction of recombinant plasmid pET21d-HtsA and expression and purification of HtsA protein
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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.
作者 宋英莉 吕春梅 张晓兰 边淑玲 徐杰 朱宛宛 朱辉
机构地区 哈尔滨医科大学生理学教研室
出处 《哈尔滨医科大学学报》 CAS 北大核心 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)
关键词 A组链球菌 HtsA 克隆 蛋白纯化 group A streptococcus HtsA protein gene cloning protein purification
分类号 Q78 [生物学—分子生物学]
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参考文献16

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同被引文献18

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哈尔滨医科大学学报
2012年 第4期

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