在减毒伤寒杆菌CVD908疫苗株中四环素诱导表达恶性疟原虫MSP1-31片段基因

Expression of Plasmodium falciparum MSP1-31 fragment in attenuated Salmonella typhi CVD908 strain induced by tetracycline controlled promoter

目的 :用含 PL tet O启动子的 p ZE11质粒在减毒伤寒杆菌 CVD90 8疫苗株中表达恶性疟原虫 MSP1- 31片段基因。 方法 :构建受四环素诱导调控的重组质粒 p ZE11/ MSP1- 31,将该重组质粒用电穿孔转化法转化入减毒伤寒杆菌 CVD90 8/ tet R疫苗株中 ,用免疫印迹反应检测 MSP1- 31在 CVD90 8/ tet R株中的四环素诱导表达。结果 :构建了重组的 p ZE11/ MSP1- 31质粒 ,免疫印迹反应显示该 MSP1- 31基因在 CVD90 8/ tet R株中得到有效表达 ,且依赖于四环素的存在。结论 :成功地构建了由四环素诱导的、表达恶性疟原虫 MSP1- 31的减毒伤寒杆菌疫苗株。

Objective:To express MSP1-31 gene of Plasmodium falciparum in Salmonella typhi CVD908 vaccine strain using a tetracycline-controlled P LtetO promoter. Methods:The MSP1-31 gene was cloned into the plasmid of pZE11 and transformed into the CVD908/tetR strain by electroporation. Expression of MSP1-31 in CVD908/tetR strain was detected using the method of Western blot. Results: The recombinant plasmid of pZE11/MSP1-31 was constructed, there was effective expression of MSP1-31 protein in CVD908/tetR strain in presence of tetracycline, and no expression of gene in absence of tetracycline. Conclusion: The recombinant Salmonella typhi strain in which the expression of Plasmodium falciparum MSP1-31 fragment induced by tetracycline is established successfully.