日本血吸虫中国大陆株22.6kD膜相关蛋白基因的克隆及其在大肠杆菌中的高效表达

Cloning of Schistosoma japonicum Chinese Strain 22.6kD Membrane associated Protein (Sj 22.6) Gene and Its Overproduction in Escherichia coli *

以日本血吸虫ｍＲＮＡ为模板，用ＲＴＰＣＲ法快速克隆到一大小约６００ｂｐ的ＤＮＡ片段，ＤＮＡ序列分析证实，所扩增到的ＤＮＡ片段中含有日本血吸虫２２６膜相关蛋白（Ｓｊ２２６（Ｃｈ））基因，将该基因重组到表达型质粒ｐＧＥＸ４Ｔ中，表达的ＧＳＴ融合蛋白分子量约４８ｋＤ，用谷胱甘肽琼脂糖凝胶亲和层析柱纯化的重组蛋白不仅纯度好，而且得率高，纯化产量可达４０ｍｇ／Ｌ培养物，免疫试验结果表明该重组蛋白具有良好的抗原性，为其在血吸虫病抗感染中的免疫作用研究创造了条件。

A 576bp DNA fragment was amplificated by RT PCR,from Schistosoma japonicum adult worm mRNA.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 48kD.The yield of expression was around 40mg/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.