检验医学与临床2016，Vol.13Issue(6)：730-733,4.DOI:10.3969/j.issn.1672-9455.2016.06.004
人胱抑素C的原核表达及活性鉴定
Prokaryotic expression and activity identification of human cystatin C
摘要
Abstract
Objective To establish a prokaryotic expression system of cystatin C (Cys C) protein for express-ing ,purifying and identifying recombinant human Cys C protein .Methods Under the premise of unchanging amino acid sequence ,the Cys C coding gene was performed the Escherichia(E .) coli synonym codon bias optimization and its sequence was artificially synthesized .The Cys C gene was amplified by PCR .The human recombinant Cys C pro-tein was expressed in the prokaryotic system .The expressed Cys C recombinant protein was purified .Its reaction sen-sitivity and immunogenicity were assayed by indirect ELISA .Results The SDS-PAGE identification indicated the re-combinant gene gained the effective expression in E .coli ,the molecular weight of recombinant Cys C protein was 35 × 103 .The Western blot results further revealed that the recombinant protein was Cys C protein ,the indirect ELISA results displayed that when the coating concentration was 0 .5 mg/L ,the positive value would be produced .Conclusion The pro-karyotic expression system is successfully established ,Cys C protein as antigen has better immunoreactivity and im-munogenicity ,which lays a foundation for preparing corresponding antigen diagnostic monoclonal antibody ,meanwhile also provides the optional raw material for developing Cys C rapid diagnosis ELISA reagent kit .关键词
人胱抑素 C/酶联免疫吸附试验/免疫反应性/免疫原性Key words
human cystain C/enzyme linked immunosorbent assay/immunoreactivity/immunogenicity引用本文复制引用
李原,刘如石,蒋琰,李桑,郑姣,何赛,吴意..人胱抑素C的原核表达及活性鉴定[J].检验医学与临床,2016,13(6):730-733,4.基金项目
湖南省科技计划项目（2014SK3100）。 （）
