淋球菌膜抗原CppB基因的克隆及在大肠杆菌和减毒鼠伤寒沙门菌中的表达

Cloning of the Gonococcal CppB Antigen Gene and its Expression in E. coli and Attenuated S.typhimurium

将PCR扩增的淋球菌CppB膜蛋白基因区段,分别克隆在游离蛋白表达载体pKK223-3和融合蛋白表达载体pWR450-1上,以点杂交证实了克隆子。控制合适表达条件,在大肠杆菌中获高效表达,免疫印迹证实其可被淋病患者血清特异识别,将融合蛋白表达质粒转入减毒鼠伤寒沙门氏菌中也获表达,为淋病口服活疫苗的制备奠定了基础。

The gene of a specific antigen protein CppB from 4. 2kb crypid plasmid of Neisseria gon-orrhoeae was cloned by PCR and gene recombinant techniques. Using pure-protein-expressing vector pKK223-3 and fused-protein-expressing vector pWR450-1,the CppB gene was highly expressed in E. coli. Western blot demonstrated its immunological reaction with sera from gonorrhoea patients. Then the fused-protein-expressing plasmid pWRJD was transformed into attenuated Salmonella systems, in which the expression of the fused-protein was also detected. The results might have brought the appearance of gonococal oral live vaccines a step nearer.