恶性疟原虫CSP基因在毕赤酵母中的分泌表达  

The gene of plasmodium falciparum circumsporozoite secretion and expression in pichia pastoris

作　　者：雷清[1] 梁凌宇[1] 王爱秀[1] 陈俏丽[1] 李刚[1] 陈勇[1] 蒋琳[1] 

机构地区：[1]兰州生物制品研究所有限责任公司第四研究室甘肃疫苗工程技术研究中心,甘肃兰州730046

出　　处：《微生物学免疫学进展》2016年第1期14-17,共4页Progress In Microbiology and Immunology

基　　金：2014年甘肃省自然科学基金(项目编号:145RJZA231)

摘　　要：目的用毕赤酵母表达系统表达恶性疟原虫环子孢子蛋白（CSP）,便于恶性疟疾疫苗的进一步研究。方法根据Gen Bank得到恶性疟原虫7G8的基因序列,选取该序列628~1 194位基因,以密码子最佳化为原则合成基因后构建酵母表达载体CSP/p GAPZa A,电转毕赤酵母菌PDI-GS115,获得酵母重组体。三角瓶规模表达重组菌,获得表达上清。结果经SDS-PAGE电泳、Western blot检测表达上清,均显示有目的蛋白表达。结论在毕赤酵母中实现了CSP基因的表达,为恶性疟疾疫苗的研发作了必要的补充。Objective To express Plasmodium falciparum circumsporozoite protein in Pichia yeast expression system so as to further study for development of malaria vaccine. Methods The sequence 628-1 194 allele was selected as the target gene according to the sequence of the published circumsporozoite protein genes of 7G8strain（ Gene Bank accession AB121015. 1）. After transformed to Pichia. pastoris PDI / GS115 strain by electroporation method,the synthetic codon-optimized gene was constructed and recombinant plasmid CSP / p GAPZa A was obtained. The positive yeast was expressed on a flask scale and supernatant was harvested. Results The expression product of the target gene was detected and identified by SDS-PAGE and Western blot. Conclusion Circumsporozoite protein gene was expressed in Pichia pastoris in the supernatant,on which an essential base was supplied for the development of malaria vaccines.

关 键 词：环子孢子蛋白 毕赤酵母 基因表达 

分 类 号：R392[医药卫生—免疫学]