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

doi:10.16431/j.cnki.1671-7236.2016.07.004

Abstract

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.According 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 constructed recombinant plasmid pMD20-T-LpxB was transformed into E.coli DH5α.The recombinant 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 prokaryotic 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 sequence of amino acids encoded LpxB gene.The results showed that the CDS of LpxB was successfully cloned and expressed.The secondary structure of LpxB protein consisted structure α -helix,extended strand,β-turn and random coil which accounted for 52.41％,14.94％,8.10％ and 24.55％,respectively.

Key words: Brucella melitensis; LpxB gene; cloning; prokaryotic expression; bioinformatics analysis

CLC Number:

NIE Xin, ZHAO Tian-jing, CAO Rui-yong, PENG Dong-mei, LI Guo-hua, LI Ya-ying, XU Kai-lian, ZHU Hua-pei, PANG Feng, WANG Feng-yang, DU Li. Cloning, Prokaryotic Expression and Bioinformatics Analysis of LpxB Gene of Brucella melitensis[J]. , 2016, 43(7): 1681-1687.

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Cloning, Prokaryotic Expression and Bioinformatics Analysis of LpxB Gene of Brucella melitensis

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