PSCA-HSP70融合蛋白在大肠杆菌中的表达与纯化  

Expression and Purification of Recombinant PSCA-HSP70 Protein in E.coli

作　　者：董磊[1,2] 张晓鹏[1] 房婷[1] 易绍琼[1] 于婷[1] 侯利华[1] 付玲[1] 于长明[1] 陈薇[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [2]空军总医院临床检验中心,北京100142

出　　处：《生物技术通报》2011年第6期150-154,共5页Biotechnology Bulletin

摘　　要：旨在大肠杆菌中表达PSCA-HSP70融合蛋白,并对其进行纯化。克隆人PSCA基因及HSP70基因,构建表达PSCA-HSP融合蛋白的工程菌,优化表达及纯化条件,对重组蛋白进行纯化。结果表明,成功构建重组表达质粒PSCA-HSP,重组蛋白得到可溶性表达,优化纯化条件后获得90%以上纯度的重组蛋白。本研究成功实现了PSCA与HSP的融合表达,为下一步肿瘤疫苗的研制奠定基础。The study was to construct recombinant plasmid and express the PSCA-HSP fusion protein in E. coli. The PSCA and HSP70 genes were amplified by PCR,then PSCA gene was cloned into prokaryote expression vector pET-21a（ ＋ ）with HSP70 gene af- ter digestion with Nde I,EcoR I and Xho I to construct recombinant plasmid pET21a-PSCA-HSP, pET21a-PSCA-HSP was expressed in E. coli and detected with Western blotting. The recombinant protein was purified with optimization of expression and purification. SDS- PAGE was used to analyze the purity of the purified recombinant protein. Results showed that the recombinant plasmid pET21a-PSCA- HSP was successfully constructed and the PSCA-HSP was successfully expressed in soluble form in E. coll. After purification,the purity of recombinant protein reached above 90%. The recombinant protein PSCA-HSP was successfully expressed in soluble condition, which lays the foundation for development of vaccine for prostate cancer.

关 键 词：前列腺干细胞抗原 热休克蛋白 表达纯化 

分 类 号：R378[医药卫生—病原生物学]