吉林农业大学学报Issue(6)：701-706,6.DOI:10.13327/j.jjlau.2014.2169
结核分枝杆菌酯酶Rv1400c原核表达及表达产物的酶活性分析∗
Prokaryotic Expression of Esterase Rv1400c from Mycobacterium tu-berculosis and Determination of Expressed Product Enzyme Activity
摘要
Abstract
This study aims to explore the activity of esterase Rv1400c of Mycobacterium tuberculosis and do further researches on the structure and function of Rv1400c. The Rv1400c gene was ampli⁃fied with PCR and cloned into pET-28b(+) prokaryotic expression vector by using Mycobacterium tuberculosis H37Rv genomic DNA as template. Recombinant plasmid of pET28b-Rv1400c was con⁃structed and then translated into BL21 ( DE3 ) expression strain. It was induced to express with IPTG after bacterial splitting and then purified with Ni-NTA affinity chromatography method. Activi⁃ ty of the purified Rv1400c was analyzed with different substrates, pH and temperature of three direc⁃tions. The results showed that recombinant plasmid of pET28b-Rv1400c was successfully construc⁃ted;SDS-PAGE and Western blot showed Rv1400c was expressed in inclusion bodies and molecu⁃lar weight of the protein was 39 ku;The Rv1400c was found active within the range of C2 to C14 in the screening of eight kinds of substrates and the best activity was in C2;Esterase activity was the highest under the conditions of pH 8�0 and 37 ℃ when the C12 was the substrate.关键词
结核分枝杆菌酯酶Rv1400c/pET28b-Rv1400c重组质粒/包涵体/酯酶活性Key words
esterase Rv1400c of Mycobacterium tuberculosis/recombinant plasmid of pET28b-Rv1400c/inclusion body/esterase activity分类
农业科技引用本文复制引用
陶荣珊,李金伟,刘思国,王全凯..结核分枝杆菌酯酶Rv1400c原核表达及表达产物的酶活性分析∗[J].吉林农业大学学报,2014,(6):701-706,6.基金项目
国家国际科技合作专项（2011DFA32900），国家自然科学基金项目（31100659，31272538），国家重点基础研究发展计划项目［973计划（2012CB518801）］，长春市科技局项目［长科技合（2011239）］ （）
