重组人抗缪勒管激素C-末端蛋白的原核表达及纯化  

Prokaryotic expression and purification of recombinant C-terminal protein of human anti-Müllerian hormone

作　　者：林丽淑 龙韵洪 徐丽惠 冯振通 谢陈莹 陈睿 王革非[2] LIN Li-shu;LONG Yun-hong;XU Li-hui;FENG Zheng-tong;XIE Chen-ying;CHEN Rui;WANG Ge-fei(Department of Clinical Laboratory,Second People′s Hospital of Futian District,Shenzhen City,Shenzhen 518049,Guangdong Province,China;不详)

机构地区：[1]深圳市福田区第二人民医院检验科,广东深圳518049 [2]汕头大学医学院微生物与免疫学考研室,广东汕头515041

出　　处：《中国生物制品学杂志》2020年第8期890-893,共4页Chinese Journal of Biologicals

基　　金：广东省科技创新战略专项基金(2018A030313548)。

摘　　要：目的原核表达重组人抗缪勒管激素(human anti-Müllerian hormone,hAMH)C-末端蛋白,并进行纯化。方法PCR法扩增hAMH C-末端蛋白基因,插入至载体pET-28a,构建重组表达质粒pET-28a-hAMH C。重组质粒转化感受态E.coli BL21(DE3),终浓度为1.0 mmol/L的IPTG诱导表达h AMH C-末端蛋白。扩大培养后进行Ni亲和层析柱纯化,12%SDS-PAGE分析纯化产物。结果经菌落PCR及测序鉴定,重组质粒pET-28a-hAMH C构建正确。表达及纯化产物均可见相对分子质量约12500的目的蛋白条带,纯度达90%以上。结论原核表达了重组hAMH C-末端蛋白,纯化后获得了较高纯度的目的蛋白。Objective To express the C-terminal protein of human anti-Müllerian hormone(hAMH)in prokaryotic cells and purify the expressed product.Methods The gene encoding C-terminal protein of hAMH was amplified by PCR and inserted into vector pET-28 a.The constructed recombinant plasmid p ET-28 a-hAMH C was transformed to E.coli BL21(DE3)and induced with IPTG at a final concentration of 1.0 mmol/L.The expressed protein was subjected to expanded culture,then purified by Ni affinity chromatography and analyzed by 12%SDS-PAGE.Results Colony PCR and sequencing proved that recombinant plasmid pET-28 a-hAMH C was constructed correctly.Target protein band with a relative molecular mass of about 12500 was observed in both expressed and purified proteins,of which the purity reached more than 90%.Conclusion Recombinant hAMH C-terminal protein was expressed in prokaryotic cells,which reached a high purity after purification.

关 键 词：人抗缪勒管激素 原核表达 基因重组 融合蛋白 Ni亲和层析纯化 

分 类 号：R34[医药卫生—基础医学]