大肠杆菌不耐热肠毒素B亚单位融合表达载体的构建及应用  

Construction and application of a fusion expression vector containing subunit B of Escherichia coli heat-labile enterotoxin

作　　者：鲁东水[1] 毛旭虎[1] 吴超[1] 邹全明[1] 

机构地区：[1]第三军医大学医学检验系临床微生物学及免疫学教研室,重庆400038

出　　处：《第三军医大学学报》2003年第14期1275-1277,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 10 0 16 1)

摘　　要：目的　构建大肠杆菌不耐热肠毒素B亚单位的融合表达载体。方法　PCR扩增LTB基因 ,并在其 3′端去掉终止密码子 ,引入编码TAPI接头 ,通过EcoRⅠ和EcoRⅤ酶切位点重组于PinpointXa Ⅰ载体上 ,将幽门螺杆菌尿素酶B亚单位与之融合 ,转化E .coliJM10 9,SDS PAGE及Westernblotting分析其表达。结果　成功构建了LTB融合表达质粒 ,并获得了幽门螺杆菌尿素酶B亚单位与之融合表达。结论　LTB融合表达载体的成功构建为研究分子内佐剂疫苗奠定了实验基础。Objective To construct a fusion expression vector with subunit B of Escherichia coli heat labile enterotoxin. Methods Gene encoding LTB without stop codon obtained by PCR was introduced to a linker and was recombined on vector Pinpoint Xa I through Eco RⅠ and Eco RⅤ sites and then fused with ureB. The recombinant was used to transform E. coli JM109. The expression of LTB was analyzed by SDS PAGE and Western blotting. Results The fusion expression plasmid was successfully constructed. The subunit B of urease of Helicobacter pylori (Hp) fusogenic protein with LTB with the ability to bind GM 1 and the reactogenicity with polyclonal antibodies against Hp was harvested. Conclusion The successfully constructed vector provides experimental base for the studies of intramolecular adjuvant vaccine.

关 键 词：不耐热肠毒素B亚单位 融合表达 载体 幽门螺杆菌尿素酶B亚单位 

分 类 号：R378.21[医药卫生—病原生物学] R394-33[医药卫生—基础医学]