日本血吸虫谷胱甘肽-S-转移酶基因和产毒大肠杆菌纤毛抗原基因的联合表达  

ASSOCIATION EXPRESSION OF GENES ENCODING GST OF SCHISTOSOMA JAPONICUM AND ENTEROTOXIGENIC ESCHERICHIA COLI *

作　　者：张威[1,2,3] 张景六[1,2,3] 刘述先[1,2,3] 

机构地区：[1]中国预防医学科学院寄生虫病研究所 [2]中国科学院上海植物生理研究所 [3]世界卫生组织疟疾,血吸虫病和丝虫病合作中心

出　　处：《中国寄生虫学与寄生虫病杂志》1999年第2期97-100,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家863高技术计划项目

摘　　要：目的：编码日本血吸虫２６ｋＤａＧＳＴ和产毒大肠杆菌纤毛抗原Ｋ９９基因的联合表达。方法：ＰＣＲ扩增的Ｋ９９基因克隆入ｐＵＣ１８载体，再将限制性酶切获得的ＧＳＴ基因克隆入ｐＵＣ１８－Ｋ９９重组质粒。限制性酶切和琼脂糖凝胶电泳选择连接方向正确的重组质粒用于重组蛋白的表达。共表达产物采用ＳＤＳ－ＰＡＧＥ、Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ、反向ＩＨＡ、ＥＬＩＳＡ和透视电镜观察进行分析鉴定。结果：共表达产物的分子量约５０ｋＤａ，存在于宿主菌体表面的纤毛中，ＧＳＴ抗血清和Ｋ９９抗血清均可识别共表达产物，提示共表达产物既有ＧＳＴ表位，又有Ｋ９９抗原表位。结论：日本血吸虫２６ｋＤａＧＳＴ和产毒大肠杆菌Ｋ９９基因共表达获得成功。AIM: To study the association expression of two genes encoding GST of Schistosoma japonicum and K99 of enterotoxigenic Escherichia coli. METHODS: PCR technique was used to gain the K99 gene. After digestion with BamHⅠ and EcoRⅠ, the gene was cloned into the plasmid vector pUC18 by using recombinant DNA techniques. The other target gene of GST in pSj5 plasmid was obtained by EcoRⅠ digestion, and was then ligated into the recombinant plasmid pUC18 K99. The expressed product was assayed by SDS PAGE, Western blotting, reversal IHA, ELISA and transmission electron microscopy. RESULTS: Co expression product of around 50 kDa was obtained. The protein was recognized by anti GST and anti K99 antibodies (ELISA and IHA), and acted as pilus on the surface of transformed DH5α E.coli bacteria. CONCLUSION: Co expression of the genes encoding GST and K99 was successfully achieved.

关 键 词：日本血吸虫 纤毛抗原 基因表达 GST 

分 类 号：R383.24[医药卫生—医学寄生虫学]