日本血吸虫脂肪酸结合蛋白基因的克隆及其在大肠杆菌中的表达  

Cloning of Schistosoma japonicum Chinese Strain Fatty Acid Binding Protein (SjFABPc) Gene and Its Overproduction in Escherichia coli

作　　者：蔡学忠[1,2] 田锷[1,2] 杨冠珍 施福恢[1,2] 沈纬[1,2] 蔡幼民 吴祥甫[1,2] 

机构地区：[1]中国农业科学院上海家畜寄生虫病研究所 [2]中国科学院上海生物化学研究所

出　　处：《生物化学与生物物理学报》1998年第2期203-206,共4页

基　　金：国家863生物高技术项目;总理专项基金

摘　　要：用ＰＣＲ法从日本血吸虫中国大陆株成虫ｃＤＮＡ文库中快速克隆出一大小为６００ｂｐ左右的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脂肪酸结合蛋白（Ｓｊ－１４ＦＡＢＰｃ）基因，将该基因重组到表达型质粒ｐＧＥＸ－２Ｔ中，表达的ＧＳＴ融合蛋白分子量是４１ｋＤ，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２５ｍｇ／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，为其用作疫苗分子创造了条件。A 600 bp DNA fragment was amplificated by PCR, from an adult Schistosoma japonicum cDNA library. Sequence analysis revealed that this fragment contained the S. japonicum Chinese Mainland strain fatty acid binding protein(Sj14FABPc) gene. This gene was then cloned into the expression vector pGEX2T,and subsequently expressed in Escherichia coli. The recombinant GSTfusion protein could be purified by glutathione agarose affinity chromatography. Its molecular weight was about 41 kD. The yield of expression was around 25 mg/L E.coli culture. The immunological test suggested that the recombinant protein had good antigenicity,and could be developed into a new vaccine molecule of S. japonicum.

关 键 词：脂肪酸结合蛋白 日本血吸虫 基因克隆 基因表达 

分 类 号：R383.24[医药卫生—医学寄生虫学] Q513[医药卫生—基础医学]