西安交通大学学报（医学版）2017，Vol.38Issue(4)：502-506,5.DOI:10.7652/jdyxb201704007
重组人细胞角蛋白9cDNA的原核表达及纯化
Prokaryotic expression and purification of the cDNA of recombinant human cytokeratin 9
摘要
Abstract
Objective To clone and fuse the cDNA of human cytokeratin 9 in prokaryotic expression system,and purify and identify the fusion protein.Methods The cDNA fragment of human cytokeratin 9 was amplified from human keratinocyte (HaCaT) total RNA with specific primers.The PCR products were cloned into vector pET-28a,then the fusion protein of his-CK9 was induced by IPTG.The expressed fusion protein of his-CK9 was purified by nickel ion affinity chromatography and identified by SDS-PAGE and Western blot.Results The sequencing proved that the recombinant vector of the cDNA of CK9 was correct.The fusion protein of his-CK9 was induced to be expressed in E.coli.The fusion protein of his-CK9 was highly purified and his-CK9 showed specific binding to the commercialized antibodies of CK9.Conclusion The recombinant vector of pET-28a-CK9 has been successfully constructed,and the fusion protein of his-CK9 has been successfully expressed and purified.关键词
原核表达系统/细胞角蛋白9/融合蛋白/细胞骨架Key words
prokaryotic expression system/cytokeratin 9/fusion protein/cytoskeleton分类
医药卫生引用本文复制引用
王博,周艳,侯卫坤,蔡永松,张英,王林玉,韩燕,孟列素..重组人细胞角蛋白9cDNA的原核表达及纯化[J].西安交通大学学报（医学版）,2017,38(4):502-506,5.基金项目
国家自然科学基金资助项目(No.81273211,81201373) （No.81273211,81201373）
陕西省社会发展科技攻关项目(No.2016SF-238) （No.2016SF-238）
西安交通大学第一附属医院基金资助项目(No.2016QN-13) Supported by the National Natural Science Foundation of China (No.81273211 and 81201373),the Scientific and Technological Project of Social Development Foundation of Shaanxi Province (No-2016SF-238),and Science Foundation of the First Affiliated Hospital of Xi'an Jiaotong University (No.2016QN-13) （No.2016QN-13）
