Cai X, Lin J, Yang G, Shi F, Shen W, Cai Y, Wu X
Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Sciences, China.
Chin J Biotechnol. 1999;15(1):7-13.
A 567bp DNA fragment was amplified from Schistosoma japonicum adult worm mRNA by RT-PCR. Sequence analysis revealed that this fragment contained S. Japonicum Chinese strain membrane-associated protein (Sj-22.6) gene. Then this gene was cloned into the expression vector pGEX-4T, and subsequently expressed in Escherichia coli. The recombinant GST-fusion protein was purified by glutathione agarose affinity chromatography. Its molecular weight was about 48 kD. The yield of expression was around 40 mg/L E.coli culture. The immunological test suggested that the recombinant protein had good antigenity which could make a good basis for the research of its immunological function in Schistosomiasis.
通过逆转录聚合酶链反应（RT-PCR）从日本血吸虫成虫mRNA中扩增出一段567bp的DNA片段。序列分析表明，该片段包含日本血吸虫中国株膜相关蛋白（Sj-22.6）基因。然后将该基因克隆到表达载体pGEX-4T中，随后在大肠杆菌中表达。重组GST融合蛋白通过谷胱甘肽琼脂糖亲和层析进行纯化。其分子量约为48kD。表达产量约为40mg/L大肠杆菌培养物。免疫学检测表明，该重组蛋白具有良好的抗原性，可为研究其在血吸虫病中的免疫功能奠定良好基础。
