摘要
目的 构建大肠杆菌不耐热肠毒素B亚单位的包涵体形式融合表达载体。方法 PCR扩增LtB基因 ,并在其 5’端去掉信号肽 ,3’端去掉终止子 ,引入编码YPQD接头 ,通过PstI +BamHI酶切位点重组于PinPointTM Xa Ⅲ载体上 ,转化E .coliJM1 0 9,SDS PAGE及Western blot分析其表达。结果 成功构建了LTB融合表达质粒 ,并将幽门螺杆菌热休克蛋白A亚单位与之融合获得了表达。结论 LTB融合表达载体的成功构建为研究分子内粘膜佐剂特性奠定了基础。
Objective To construct a fusion expression vector of Escherichia coli heat labile enterotoxin B subunit. Methods Gene encoding LtB without signal peptide and stop codon was added to the linker TAPI by PCR. It was cloned into PinPoint TM Xa Ⅲ with Pst Ⅰ and BamH Ⅰ sites. The recombinant plasmid was transfected in E.coli JM109. The expression of LTB was analysed by SDS PAGE and Western blot. Results The fusion expression plasmid was successfully constructed. The subunit A of heat shock protein of Helicobacter pylori was fused with LTB and expressed, and display both the ability of binding GM 1 and the immunogenicity with polyclonal antibodies against H. Pylori . Conclusion The vector founds a basis for the study of intramolecular adjuvant.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2001年第6期644-646,共3页
Journal of Third Military Medical University
基金
国家"九五"重点科技攻关课题 (96 90 1 0 1 5 4)
全军"九五"医药卫生科研基金资助项目 (98D0 0 4)
