羊源类鼻疽伯克霍尔德菌BPSL1467基因克隆、表达及其蛋白生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.09.007

Abstract

Abstract:

This study was aimed to clone and express BPSL1467 gene of Burkholderia pseudomallei (B.pseudomallea),and performe bioinformatics analysis of its protein.A pair of primers was designed according to the BPSL1467 gene sequence of B.pseudomallea K96243 strain in GenBank.BPSL1467 gene fragment of B.pseudomallea BPHN1 strain was amplified by PCR amplification.The BPSL1467 gene fragment was ligated into the pET-28a(+) vector to construct the pET-28a(+)-BPSL1467 recombinant plasmid.Then,the pET-28a(+)-BPSL1467 was transformed into E.coli BL21 (DE3) competent cells.The expressed product induced by IPTG was analyzed by SDS-PAGE and Western blotting.Bioinformatics analysis of BPSL1467 gene sequence was carried out using DNAMAN,ProtParam,SOPMA and Protscale softwares.The results showed that BPSL1467 gene was cloned with the length of 462 bp.The expressed recombinant protein was about 22 ku and was mainly in the form of inclusion body;The molecular weight of the BPSL1467 recombinant protein was 16 890.58 u (C₇₆₃H₁₂₀₉N₂₀₃O₂₁₇S₆);Its instability coefficient,theoretical isoelectric point (pI) and total average hydrophobicity (GRAVY) were 33.95,8.85 and -0.190 respectively,which was a stable hydrophilic basic protein;The secondary structure of the protein was mainly random coil (42.48%) and alpha helix (32.68%).This results provided a reference for the further study of molecular mechanism of Burkholderia pseudomallei BPSL1467 gene.

Key words: Burkholderia pseudomallei; BPSL1467 gene; cloning; prokaryotic expression; bioinformatics analysis

CLC Number:

S858.26

ZHANG Mengmeng, LI Baobao, CAO Ruiyong, HUANG Haifeng, ZHANG Zhenxing, YANG Xiaojian, NIE Xin, ZHU Shu, WANG Fengyang, DU Li. Cloning, Expression and Protein Bioinformatics Analysis of BPSL1467 Gene of Burkholderia pseudomallei[J]. , 2018, 45(9): 2401-2408.

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Cloning, Expression and Protein Bioinformatics Analysis of BPSL1467 Gene of Burkholderia pseudomallei

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