肾炎致病原重组受体相关蛋白的表达及纯化

用pGEX载体系统体外构建了Heymann肾炎致病原受体相关蛋白(RAP)重组表达质粒，经IPTG诱导，该质粒表达的融合蛋白在大肠杆菌中得到了高效表达，其表达量达39．4％，经GST-Sephrose 4B亲和层析，得到了高度纯化，其诱导产生的抗体经蛋白质印迹法分析证明能识别肾皮质天然抗原44ku受体相关蛋白.RAP表达及纯化的成功为研究致病原病理性表型提供了有利条件．

Expression and Purification of Recombinant Receptor-associated Protein (RAP)of the Heymann Nephritis Pathotigenic Epitope

A recombinant expression plasmid was constructed by inserting the cDNA encoding mature receptor-associated protein (RAP) into pGEX vector. High level intracellular expression of the RAP gene was Observed in the DH5a bacteria transformed with the recombinant pGEX vector after IPTG induction. The abundant GST fusion protein constituted 39. 4 % the total cellular protein. The fusion proteins were purified from bacterial lysates by using GST-Sepharose 4B affinity chromatography. The Western blot analysis showed that the rabbit anti-RAP antiserum recognized an apparent mass of 44 ku band from rat kidney microvillar protein. The high level expression and rapid purification of RAP-GST fusions was discussed.