抗菌肽CM4与人可溶性B淋巴细胞因子hsBAFF在大肠杆菌中的融合表达  

Expression of Antibacterial Peptide CM4 in Escherichia coli Fused with Human Soluble B Lymphocyte Stimulator Activing Factor

作　　者：刘海峰[1] 王小花[2] 刘平[1] 张双全[1] 

机构地区：[1]南京师范大学生命科学学院江苏省分子医学生物技术重点实验室,南京210046 [2]山西师范大学生命科学学院细胞工程研究所,临汾041004

出　　处：《中国生物工程杂志》2008年第3期32-37,共6页China Biotechnology

基　　金：国家自然科学基金(30271093);江苏省自然科学基金(BK2005140)资助项目

摘　　要：为进一步探讨抗菌肽CM4的原核表达及其生物学功能,研究了抗菌肽CM4与人可溶性B淋巴细胞刺激因子hsBAFF的融合表达及抗菌肽CM4的生物学活性。运用PCR把B淋巴细胞因子hsBAFF和家蚕抗菌肽CM4进行基因融合,构建了融合表达载体pET28a(+)/CM4-hsBAFF,并在大肠杆菌中获得高可溶性表达的融合靶蛋白,且存在于超声破碎后的上清,经分子筛Sephadex G-75纯化后的重组融合蛋白用SDS-PAGE和Western blot分析鉴定.SDS-PAGE分析表明:可以通过分子筛一步纯化得到融合蛋白,该重组融合蛋白的分子量约22.0kDa。Western blot结果显示该重组蛋白能与鼠抗人hsBAFF的抗体发生特异性反应.运用基因工程的方法获得CM4-hsBAFF重组融合蛋白,并具有很好的抑菌生物学活性。To explore the expression and function of antibacterial peptide CM4, the expression and biological activity of recombinant fusion protein CM4 - hsBAFF were studied. The human soluble B lymphocyte stimulator activing factor （hsBAFF） gene was fused to the sequence encoding CM4 to construct an expression vector pET28a （ ＋ ）/CM4 - hsBAFF. The recombinant protein was high expression of soluble recombinant protein in Escherichia coli cells, and existed in the supernatant after sonication. Recombinant fusion protein which was purified through size-exclusion chromatography was identified by SDS-PAGE and Western blot analysis. SDS-PAGE and Western blot indicated that recombinant protein was secreted as a protein of around 22.0kDa2 and can be specially recognized anti-hsBAFF antibody. The recombinant protein can be expressed and displays antimicrobial activity.

关 键 词：抗菌肽CM4 人可溶性B淋巴细胞因子 表达 纯化 

分 类 号：R392.11[医药卫生—免疫学]