大肠杆菌不耐热肠毒素B亚单位在干酪乳杆菌中的分泌表达  

E. coli Heat-labile Enterotoxin B Subunit Secretory Expression in Lactobacillus Casei

作　　者：付锦楠[1] 杨罡[1] 何顺华[1] 龚小华[1] 郭建军[1] 

机构地区：[1]江西省科学院微生物研究所,江西南昌330096

出　　处：《江西科学》2013年第6期764-767,共4页Jiangxi Science

基　　金：江西省科技支撑计划(20122BBG70093);江西省青年科学基金(2013BAB214014);江西省科学院项目(2012-YQC-08)

摘　　要：以大肠杆菌和乳酸菌穿梭表达型载体pMG36e为基本模型,采用重叠延伸PCR方法拼接相关调控元件构建一种乳酸杆菌组成型分泌表达载体;将编码LTB蛋白的eltb基因片段插入干酪乳杆菌分泌型表达载体中,构建重组质粒pMG36e-UP/L-LTB,电转化于干酪乳杆菌393中,筛选获得重组干酪乳杆菌,应用western blot方法鉴定LTB表达情况。Western-blot分析显示,约有12 KDa的目的蛋白以分泌形式表达,可被LTB阳性血清识别。表明LTB蛋白在重组干酪乳杆菌获得了表达,并且具有免疫反应活性和佐剂活性,为以LTB为分子佐剂研制乳酸菌黏膜疫苗奠定基础。pMG36e is a shuttle vector suitiable for cloning in both E. coli and Lactobacillus. The splicing primers of regulation element constructed a type of expression vector of lactic acid bacteria, the eltb gene was cloned into the expression vector, and transformed into L. casei 393 by electroporation. LTB protein expression was detected by western blot and indirect ELISA. The results showed that the LTB protein was expressed and secreted into the culture meduim. The results showed that a molecular weight of about 12kDa was expressed and secreted into the culture meduim,and the result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by anti-LTB serum. The approach described in this study will enable the efficient production of a non-toxic expressed enterotoxin as a vaccine against the toxin itself or as a carrier or adjuvant for o- ral vaccine delivery in future.

关 键 词：不耐热肠毒素B亚单位 干酪乳杆菌 分泌表达 

分 类 号：Q78[生物学—分子生物学]