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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 inclusion body of Escherichia coli heat-labile enterotoxin B subunit. Methods The LtB gene was amplified by PCR. The signal peptide was deleted at the 5 ’end and the terminator was deleted at the 3’ end. The YPQD linker was introduced into the vector, and then inserted into PinPointTM Xa Ⅲ vector by PstI + BamHI restriction enzyme digestion to transform E. coli JM10 9, SDS PAGE and Western blot analysis of its expression. Results The LTB fusion expression plasmid was successfully constructed and the Helicobacter pylori heat shock protein A subunit was fused with it. Conclusion The successful construction of LTB fusion expression vector lays the foundation for studying the characteristics of intramolecular mucosal adjuvant.