局解手术学杂志2013，Vol.22Issue(4)：349-352,4.DOI:10.11659/jjssx.1672-5042.201304002
His-AMPKα1312截短缺失融合蛋白原核表达载体的构建与纯化
Prokaryotic expression and purification of rat c-terminal truncation mutant of the AMPK fusion protein
摘要
Abstract
Objective To express rattus His-AMPKα1312 truncated fusion protein in E.coli,the expressed product for its bioactivity were purified and analyzed.Methods The truncated AMPKα1 gene was amplified by RT-PCR and inserted into prokaryotic expression vector PET28a(+).The constructed recombinant plasmid YH2/PET28a(+) was transformed to DH5α E.coli for expression under induction of IPTG.The expressed His-AMPKα1 312 fusion protein was purified by Ni2+-NTA Agarose affinity chromatography and identified for reactogenicity.Results The DNA sequence of amplified truncated AMPKαl gene was consistent with that reported in GenBank.Recombinant plasmid YH2/PET28a(+) was constructed correctly.The expressed recombinant protein,with a relative molecular mass of about 38kd,showed good reactogenicity.Conclusion The His-AMPKα1312 truncated fusion protein was successfully constructed and purified with bioactivity,which laid a foundation of further study on the function of AMPK.关键词
腺苷酸活化蛋白激酶/原核表达/蛋白纯化Key words
AMP-activated protein kinase(AMPK)/prokaryotic expression/protein purification分类
生物科学引用本文复制引用
颜洪,张琼,吴丹,宋华培..His-AMPKα1312截短缺失融合蛋白原核表达载体的构建与纯化[J].局解手术学杂志,2013,22(4):349-352,4.基金项目
国家自然科学基金面上项目(30600649) （30600649）
军队十二五重点项目(BWS11J039) （BWS11J039）
