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及其GM1结合活性分析  

E. coli heat-labile enterotoxin B subunit secretory expression in Lactobacillus casei and binding with GM1 receptor

作　　者：葛俊伟[1] 姜艳平[1] 汪淼[1] 刘敏[1] 乔薪瑗[1] 唐丽杰[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2010年第7期521-523,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"十一五"科技支撑计划(2009BADB4B01);东北农业大学博士启动基金

摘　　要：为获得表达大肠杆菌不耐热肠毒素B亚单位(LTB)的乳酸菌表达系统,并分析其免疫反应性和神经节苷脂受体(GM1)结合活性,本研究将编码LTB蛋白的eltb基因片段插入干酪乳杆菌分泌型表达载体pPG-2中,构建重组质粒pPG-2-eltb,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和间接ELISA方法鉴定LTB表达情况,并检测其与GM1结合活性。结果显示,目的蛋白以分泌形式表达,可被LTB阳性血清识别。GM1-ELISA试验结果证实表达的LTB可与牛GM1特异性结合。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。To evaluate the feasibility of recombinant Lactobacillus casei 393 expressing E.coli heat-labile enterotoxin B（LTB） subunit as an adjuvant for oral vaccine,the eltb gene was cloned into the expression vector pPG-2,and transformed into L.casei 393 by electroporation.LTB protein expression was detected by western blot and indirect ELISA.The immunogenicity and adjuvanticity of the LTB recombinant protein were determined by GM1-ELISA.The results showed that the LTB protein was expressed and secreted into the culture meduim.The recombinant protein possessed the LTB antigenic specificity and the GM1 ganglioside receptor binding activity as shown by GM1-ELISA.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 oral vaccine delivery in future.

关 键 词：LTB 干酪乳杆菌 表达 GM1结合活性 

分 类 号：S852.4[农业科学—基础兽医学]