东亚正钳蝎毒素基因BmKbpp的原核表达及其抗菌功能初步鉴定

江汉大学学报（自然科学版） ›› 2012, Vol. 40 ›› Issue (5): 64-70.

东亚正钳蝎毒素基因BmKbpp的原核表达及其抗菌功能初步鉴定

罗锋，李正，范丽梅，梁勇，林燕，蒋琳

江汉大学医学院，湖北武汉430056

收稿日期:2012-05-21 出版日期:2012-10-20 发布日期:2013-11-07
作者简介:罗锋(1975—)，男，讲师，博士，研究方向:毒素分子生物学。
基金资助:
武汉市科技局青年科技晨光计划项目(200950199019－03)；武汉市市属高等学校科学研究重点项目(20070705)；湖北省高等学校优秀中青年科技创新团队项目(T2012)

Prokaryotic Expression and Functional Characterization of Scorpion Toxins Gene BmKbpp from Buthus Martensii Karsch

LUO Feng,LI Zheng,FAN Li－mei,LIANG Yong,LIN Yan,JIANG Lin

School of Medicine,Jianghan University,Wuhan 430056,Hubei,China

Received:2012-05-21 Online:2012-10-20 Published:2013-11-07

摘要/Abstract

摘要： 一级结构分析和二级结构预测表明无二硫键蝎毒液多肽(NDBPs)BmKBPP具两亲性α－Helix结构，含有大量带净正电荷的碱性残基(Arg和Lys)，符合多价阳离子抗菌肽的典型特征。采用PCR法和质粒载体pGEX－5X－1构建BmKbpp、EGFP及BmKbpp／EGFP与GST的原核融合表达载体，3个表达载体与pGEX－5X－1分别在Rossetta(DE3)菌中进行表达，并测定4个Rossetta(DE3)表达工程菌诱导表达的生长曲线。结果表明，BmKBPP是一个具有很强抑菌功能的抗菌肽，GST、BmKBPP及EGFP3个蛋白一起融合表达可获得少量蛋白的表达，这将为后续的功能研究和应用研究奠定基础。

关键词: 蝎毒素, 抗菌肽, BmKbpp, 原核表达

Abstract: Primary structure analyzing and secondary structure predicting indicate that BmKBPP of NDBPs (Non－Disulfide－Bridged Peptides) might be a polycovalent cationic host defense peptide, which has a typical structure of amphiphilic α－helix and plenty of basic amino acids(Arg and Lys)with net positive charges. Using PCR method and plasmid vector pGEX－5X－1, the recombinant expression vectors of BmKbpp, EGFP and BmKbpp／EGFP prokaryoticly fused with GST were constructed. These three expression vectors and pGEX－5X－1 were transformed into Rossetta(DE3) strains, respectively. Expression of BmKbpp gene in E. coli demonstrates that its product has antimicrobial function by testing the living curve of host cell. Although the fusing products of genes is toxic to host cell, the slight fusion expression of GST, BmKBPP and EGFP is successful, which provides us a way to obtain BmKBPP for further functional research.

Key words: scorpion toxins, antimicrobial peptides, BmKbpp, prokaryotic expression(责任编辑:陈　旷)

中图分类号:

罗锋，李正，范丽梅，梁勇，林燕，蒋琳. 东亚正钳蝎毒素基因BmKbpp的原核表达及其抗菌功能初步鉴定[J]. 江汉大学学报（自然科学版）, 2012, 40(5): 64-70.

LUO Feng,LI Zheng,FAN Li－mei,LIANG Yong,LIN Yan,JIANG Lin. Prokaryotic Expression and Functional Characterization of Scorpion Toxins Gene BmKbpp from Buthus Martensii Karsch[J]. Journal of Jianghan University(Natural Science Edition), 2012, 40(5): 64-70.

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