摘要
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ＧＳＴＳｅｐｈｒｏｓｅ４Ｂ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．
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 RAPGST fusions was discussed.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1996年第3期254-257,共4页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金
关键词
受体相关蛋白
提纯
表达
肾炎
receptor-associated protein
Expression
purification
