重组人骨保护素片段OPG^(179)与人血清白蛋白在毕赤酵母中的融合表达  

Expression of the rhOPG^(179)-HSA Fusion Protein in Pichia pastoris

作　　者：刘俊丽[1] 刘波[2] 宋淑军[1] 司少艳[1] 谭小青[1] 吴军[2] 景青萍[1] 张建中[1] 

机构地区：[1]解放军第306医院,北京100101 [2]军事医学科学院生物工程研究所,北京100071

出　　处：《生物技术通讯》2011年第1期15-18,23,共5页Letters in Biotechnology

基　　金：总装后司令部及卫生局项目(07ZH10);306医院院级课题(07QN10)

摘　　要：目的:在毕赤酵母中融合表达人骨保护素(OPG)片段OPG179与人血清白蛋白(HSA)。方法:通过RT-PCR扩增获得OPG179基因,构建表达质粒pHILD2-rhOPG179-HSA,转化至毕赤酵母菌中进行表达、纯化,对分泌表达产物进行SDS-PAGE及Western印迹检测。结果:酶切鉴定与测序结果显示重组表达质粒pHILD2-rhOPG179-HSA正确;经Western印迹检测,融合蛋白rhOPG179-HSA的相对分子质量约为97×103,符合预期值;rhHSA-OPG179在毕赤酵母中的表达量约为80 mg/L。结论:表达并纯化制备了重组骨保护素片段与人血清白蛋白的融合蛋白,为后续的生物学活性研究奠定了一定的基础。Objective：To express in Pichia pastoris and purify recombinant human osteoprogerin（OPG） fragment OPG^179 and human serum albumin（HSA） fusion protein.Methods：The gene of OPG^179 was amplified by RT-PCR.The plasmid of pHILD2-rhOPG^179-HSA was constructed and then transformed to Pichia pastoris.The rhOPG^179-HSA fusion protein was expressed,purified,and analyzed with SDS-PAGE and Western blot.Results：The expression plasmid of pHILD2-rhOPG^179-HSA was proved correct after the plasmid was identified and sequenced.The molecular mass of fusion protein was about 97 kD.The rhOPG^179-HSA fusion protein was expressed and purified successfully from Pichia pastoris with relative higher-efficient expression level（about 80 mg/L）.Conclusion：The secreted recombinant fusion protein rhOPG^179-HSA has been successfully expressed and purified,and supply for a foundation for its biological activity analysis

关 键 词：人骨保护素 人血清白蛋白 融合蛋白 毕赤酵母 表达 纯化 

分 类 号：Q78[生物学—分子生物学]