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

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。

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

Primary structure analyzing BmKBPP of NDBPs （Non-Disulfide-Bridged and secondary structure predicting indicate that Peptides） might be a polycovalent cationic host de- fense peptide, which has a typical structure of amphiphilic a-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.