植物乳杆菌JPP2胆盐水解酶相关基因克隆和表达

Cloning and expression of bile salt hydrolase gene from Lactobacillus plantarum JPP2

通过PCR方法从植物乳杆菌JPP2中扩增出胆盐水解酶(BSH)相关基因bsh3,利用中间克隆载体pMID19-T将其构建于表达载体pET-28b上,并转化入表达宿主菌E.coli BL21(DE3),成功构建重组BSH的工程菌。核苷酸及推导的氨基酸序列分析表明,正确克隆出目的基因。诱导表达后,SDS-PAGE电泳结果显示出特异性蛋白质条带,其分子量约为38 kDa。此单克隆体系的构建为进一步研究BSH的功能奠定基础。

The bile salt hydrolase＇s gene bsh3 was amplified from Lactobacillus plantarum JPP2 by PCR technique. The purified PCR products were inserted into expression vector pET-28b by means of intermedia cloning vector pMD19-T. The positive plasmid was transformed into E. coli BL21 （DE3）, and the recombinant strain E. coli pET28-bsh3 was obtained. The results showed by SDS-PAGE analysis that bsh3 was expressed and the molecular weight was 38 kDa. The construction of recombinant bsh3 would facilitate the further research on bile salt hydrolase function in deconjugation of bile salts.