摘要
目的 用含PLtetO启动子的pZE11质粒在减毒伤寒杆菌CVD90 8疫苗株中表达恶性疟原虫主要裂殖子表面蛋白。方法 将克隆有恶性疟原虫裂殖子表面蛋白C末端基因 (MSP1 42 )的重组pZE11质粒用电穿孔转化法转化入减毒伤寒杆菌CVD90 8/tetR疫苗株中 ,用SDS PAGE、免疫印迹反应和免疫荧光反应检测MSP1 42在CVD90 8/tetR株中的体内外四环素诱导表达。结果 构建了CVD90 8/tetR/MSP1 42菌株 ;该菌株不仅在体外 ,而且在小鼠的肝脾组织中 ,由四环素诱导表达了MSP1 42蛋白 ;在体外 ,诱导表达的表达量要高于组成性的表达和tetR基因存在于质粒上的表达。结论 成功地构建了由四环素诱导的、表达恶性疟原虫MSP1 42蛋白的减毒伤寒杆菌重组菌。
Objective To investigate the inducible expression of MSP1 gene of Plasmodium falciparum in Salmonella typhi CVD908 vaccine strain using a tetracycline controlled P LtetO promoter. Methods The recombinant plasmid pZE11/MSP1 42 was transferred into the CVD908/tetR strain by electroporation. Detections of the expression of MSP1 42 both in vitro and in vivo were carried out using SDS PAGE, Western blot and immunofluorescence assay. Results The CVD908/tetR/MSP1 42 strain was constructed and the expression of MSP1 42 was dependent on the presence of tetracycline in vitro. The yield of the inducible expression was higher than that of constitutive system. Moreover, the MSP1 42 was expressed in the liver and spleen of mice inoculated with the CVD908/tetR/MSP1 42 strain in the presence of tetracycline, whereas no expression was detected in the absence of the inducer. Conclusion The recombinant Salmonella typhi strain which expresses the MSP1 42 fragment of Plasmodium falciparum induced by tetracycline has been established successfully.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2000年第10期780-783,共4页
National Medical Journal of China
基金
国家863计划(102070404)
国家自然科学基金资助项目(39780024)
关键词
沙门氏菌
伤寒
疟原虫
恶性
四环素
基因表达
Salmonella typhi
Plasmodium falciparum
Tetracycline
Gene expression
