中国畜牧兽医2016，Vol.43Issue(7)：1681-1687,7.DOI:10.16431/j.cnki.1671-7236.2016.07.004
羊种布鲁氏菌LpxB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 Bioinformatics Analysis of LpxB Gene of Brucella melitensis
摘要
Abstract
The study was aimed to clone and express LpxB gene,and perform the bioinformatics analysis of protein.The genomic DNA of Brucella melitensis M5-90 was used as template.Ac-cording to the genome sequence of M5-90 on GenBank,a pair of primers was designed.LpxB gene,which was 1 188 bp,was amplified by PCR,and was ligated into pMD20-T vector.The con-structed recombinant plasmid pMD20-T-LpxB was transformed into E.coli DH5α.The recombi-nant plasmid was confirmed by endonuclease digestion and sequencing.The coding region of LpxB from pMD20-T was digested by BamHⅠ and XhoⅠ.Then,the fragment was inserted into pro-karyotic expression vector pET-28a,and the positive plasmid was named pET-28a-LpxB.The pET-28a-LpxB was transformed into E.coli BL21 (DE3).The expressed protein was identified by SDS-PAGE and Western blotting.DNAMAN and BioEdit softwares were used to analyze the se-quence of amino acids encoded LpxB gene.The results showed that the CDS of LpxB was success-fully cloned and expressed.The secondary structure of LpxB protein consisted structureα-helix, extended strand,β-turn and random coil which accounted for 52.41%,14.94%,8.10% and 24.55%,respectively.关键词
羊种布鲁氏菌/LpxB基因/克隆/原核表达/生物信息学分析Key words
Brucellamelitensis/LpxB gene/cloning/prokaryotic expression/bioinformatics analysis分类
生物科学引用本文复制引用
聂鑫,王凤阳,杜丽,赵天靖,曹瑞勇,彭冬梅,李国华,李亚颖,徐开莲,朱华培,庞峰..羊种布鲁氏菌LpxB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,2016,43(7):1681-1687,7.基金项目
国家自然科学基金(31460670) （31460670）
