人CD19分子胞外区在大肠杆菌中的表达研究

Expression of the extracellular domain of human CD19 gene in Escherichia coli

本实验应用计算机优化设计，利用ＰＣＲ 和分子克隆技术，以ｐＵＣ１９／ＣＤ１９ 重组质粒为模板，扩增出人ＣＤ１９ 分子的胞外区基因，并与表达载体相连，经３０ ℃扩增菌体，４２ ℃诱导，实现了在Ｅ－ｃｏｌｉ 中融合表达相对分子质量（ Ｍｒ） 为４１ ０００ 的蛋白谱带。初步纯化的包涵体可溶于２ ％ ＳＤＳ，为对其单克隆抗体（ｍＡｂ） 的制备奠定了基础。

The gene fragment encoding extracellular domain of CD19 obtained by PCR was inserted into pGEM T and sequenced Then the recombinant plasmids pEX31c CD19 were cloned and expressed in E coli YK537 After induction at 42℃, a new anticipated protein band with relative molecule mass(Mr) of 41 000 appeared on SDS PAGE gel and Western blot Thus,the successful expression of CD19 gene in E coli YK537 has laid a foundation to prepare the anti CD19 mAb