按蚊TEP1基因的原核表达及鉴定

Prokaryotic expression and identification of anophele TEP1

目的原核表达斯氏按蚊TEP1蛋白。方法PCR方法从斯氏按蚊扩增TEP1基因,并插入原核表达载体pQE-80L,构建重组表达质粒pQE-80L/TEP1。转化DH5α菌,IPTG诱导表达融合蛋白及质谱鉴定。结果成功表达重组质粒,经过质谱鉴定为斯氏按蚊TEP1分子。结论成功表达了按蚊TEP1蛋白为后续的抗体制备及TEP1功能研究奠定了基础。

To obtain the Anopheles stephensi （As） TEP1,its gene was ampfified by PCR from the Anopheles stephensi. and the fragment was inserted into vector pQE-80L to construct recombinant plasmid pQE-80L/TEP1. Following transformation into E. coli DH5a, the fusion protein was expressed under IPTG induction. SDS-PAGE and mass spectrometry （MS） were used to determine the expression of the expected protein. The fusion protein was successfully expressed and the TEP1 protein was confirmed by MS. The expression of the recombinant TEP1 protein has laid a foundation for further antibody preparation and function studies of TEP1 protein.