人肿瘤抑制因子Folliculin(FLCN)蛋白的原核表达纯化

Prokaryotic Expression and Purification of Human Tumor Suppressor Folliculin(FLCN) Protein

肿瘤抑制因子Folliculin(FLCN)的突变或缺失会引起常染色体显性遗传病——BHD综合征,但其诱发机制不清,因而认知FLCN的结构和功能非常必要。本文利用优化pGEX-6p-1-GST-FLCN重组质粒构建了大肠杆菌原核表达系统,诱导表达GST-FLCN融合蛋白,再通过GST介质亲和层析和分子筛纯化,最后通过Xa因子蛋白酶切除GST标签,收集获取了高纯度(90%左右)的FLCN蛋白,为FLCN的结构与功能分子奠定了基础。

Folliculin(FLCN)is a tumor suppressor,its mutation or deletion causes autosomal dominant genetic disease BHD syndrome,but its induction mechanism is unclear.Therefore,it is necessary to recognize the structure and function of FLCN.In this paper,the prokaryotic expression system of E.coli was constructed by optimizing the recombinant plasmid pGEX-6p-1-GST-FLCN,which induced the expression of GST-FLCN fusion protein,and then purified by GST medium affinity chromatography and molecular sieve.Finally,the GST label was excised by factor Xa protease.The collection of high purity FLCN protein laid the foundation for the structural and functional molecules of FLCN.