中国畜牧兽医2018，Vol.45Issue(2)：302-309,8.DOI:10.16431/j.cnki.1671-7236.2018.02.003
类鼻疽伯克霍尔德菌groEL基因的克隆、原核表达及其蛋白的生物信息学分析
Cloning, Prokaryotic Expression and Protein Bioinformatics Analysis of groEL Gene of Burkholderia pseudomallei
摘要
Abstract
This study was aimed to clone and express groEL gene of Burkholderia pseudomallei,and perform the bioinformatics analysis of groEL protein.The genome DNA of the bacteria was extracted as the template,a pair of primers were designed with reference to the groEL gene sequence of Burkholderia pseudomallei in GenBank.The groEL gene fragment of 1 641 bp was amplified by PCR,and ligated into pMD19-T vector to construct pMD19-T-groEL recombinant plasmid.The recombinant plasmid pET-28a (+)-groEL was constructed by digestion with BamH Ⅰ and Hind Ⅲ.The correct pET-28a(+-)-groEL was transformed into E.coli BL21 (DE3) competent cells and induced by IPTG.Protein was identified by SDS-PAGE and Western blotting.DNAMAN and BioEdit softwares were used to perform the bioinformatics analysis.The results showed that groEL gene of Burkholderia pseudomallei was successfully cloned and expressed.The fusion protein was about 64 ku.The formular of GroEL protein was C4510H7381N1641O1840S521,the molecular mass was 15 893,the extinction coefficient was 32 500,the instability index was 40.31,and the average hydrophilicity was 0.901.The alpha helix (Hh),extended chain (Ee) and random coil (Cc) accounted for 48.71％,13.19％ and 38.10％,respectively.This results laid a foundation for the further study of molecular mechanism of Burkholderia pseudomallei groEL gene.关键词
类鼻疽伯克霍尔德菌/groEL基因/克隆/原核表达/蛋白质鉴定/生物信息学分析Key words
Burkholderia pseudomallei/groEL gene/cloning/prokaryotic expression/protein identification/bioinformatics analysis分类
生物科学引用本文复制引用
黄海峰,李亚颖,王凤阳,杜丽,张振兴,杨小健,曹瑞勇,李宝宝,聂鑫,朱姝,李国华,彭冬梅..类鼻疽伯克霍尔德菌groEL基因的克隆、原核表达及其蛋白的生物信息学分析[J].中国畜牧兽医,2018,45(2):302-309,8.基金项目
国家现代农业产业技术体系(CARS-38) （CARS-38）
海南省重大科技计划项目(ZDKJ2016017-01) （ZDKJ2016017-01）
