期刊论文

Prokaryotic expression and purification of the Acinetobacter baumannii DsbC protein

中文

中国病原生物学杂志

1673-5234

2020

15

02

164-167

10.13350/j.cjpb.200208

国家自然科学基金项目（No.31500156）；

本校为第一机构

目的 克隆鲍曼不动杆菌二硫键形成蛋白C(DsbC)基因,制备重组DsbC蛋白,为研究其活性奠定基础. 方法 通过NCBI获得DsbC蛋白序列,分析其生物学性质.采用PCR方法扩增无信号肽DsbC基因,并将其连接到表达质粒pET28a,构建重组质粒pET28a-DsbC,转化大肠埃希菌BL21 (DE3)后用IPTG诱导DsbC蛋白的表达,采用镍离子亲和层析法纯化目的蛋白. 结果 DsbC具有一个信号肽,由2个相同的DsbC蛋白分子构成二聚体.PCR扩增去掉信号肽的DsbC基因大小为636 bp,构建的重组质粒pET28a-DsbC经PCR测序验证正确.重组质粒转化DE3后经IPTG诱导,表达分子质量单位为26.1×103的DsbC蛋白,经镍柱纯化得到单一电泳条带的重组蛋白. 结论...

Objective To clone the disulfide bond forming protein C (DsbC) gene of Acinetobacter baumannii and prepare a recombinant DsbC protein in order to provide a basis for the study of its activity. Methods The DsbC protein sequence was obtained from NCBI, and its biological properties were analyzed. The DsbC gene without a signal peptide was amplified with PCR and ligated into the expression plasmid pET28a to construct the recombinant plasmid pET28a-DsbC. The pET28a-DsbC was transferred into E. coli BL21(DE3). Expression of the DsbC protein without ...