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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。
OBJECTIVE: To co-express the gene encoding 26kDaGST of Schistosoma japonicum and toxin-producing K99 gene of toxigenic E. coli. Methods: The K99 gene amplified by PCR was cloned into pUC18 vector, and the GST gene obtained by restriction enzyme digestion was cloned into pUC18-K99 recombinant plasmid. Restriction endonuclease and agarose gel electrophoresis to select the correct direction of the recombinant plasmid for recombinant protein expression. The co-expression products were identified by SDS-PAGE, Western blotting, reverse IHA, ELISA and transmission electron microscopy. Results: The molecular weight of the co-expression product was about 50 kDa, which existed in the cilia on the surface of the host cell. Both the GST antiserum and the K99 antiserum could recognize the co-expressed product, suggesting that the co-expressed product had both the GST epitope and the K99 epitope . Conclusion: The co-expression of Schistosoma japonicum 26 kDa GST and toxigenic E. coli K99 gene was successful.