[1]王红红,熊意,马亚茹,等.羊种布鲁氏菌043新疆流行株WbdA基因和WbkE基因的原核表达及生物信息学分析[J].江苏农业学报,2017,(02):367-372.[doi:doi:10.3969/j.issn.1000-4440.2017.02.020]
　WANG Hong-hong,XIONG Yi,MA Ya-ru,et al.Prokaryotic expression and bioinformatics analysis of WbdA gene and WbkE gene of 043 Xinjiang strains of Brucella melitensis[J].,2017,(02):367-372.[doi:doi:10.3969/j.issn.1000-4440.2017.02.020]
点击复制

羊种布鲁氏菌043新疆流行株WbdA基因和WbkE基因的原核表达及生物信息学分析()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年02期
页码:
367-372
栏目:
畜牧兽医·水产养殖
出版日期:
2017-04-30

文章信息/Info

Title:
Prokaryotic expression and bioinformatics analysis of WbdA gene and WbkE gene of 043 Xinjiang strains of Brucella melitensis
作者:
王红红1; 熊意1; 马亚茹1; 李爽2; 张辉2; 陈创夫2
（1.新疆石河子大学生命科学学院， 新疆石河子832000；2.新疆石河子大学动物科技学院， 新疆石河子832000）
Author(s):
WANG Hong-hong1; XIONG Yi1; MA Ya-ru1; LI Shuang2; ZHANG Hui2; CHEN Chuang-fu 2
( 1.College of Life Technology，Shihezi University，Shihezi 832000，China;2.College of Animal Science and Technology，Shihezi University，Shihezi 832000，China）
关键词:
布鲁氏菌; WbdA基因; WbkE基因; 原核表达; 生物信息学
Keywords:
Brucella; WbdA gene; WbkE gene; prokaryotic expression; bioinformatics
分类号:
S852.4
DOI:
doi:10.3969/j.issn.1000-4440.2017.02.020
文献标志码:
A
摘要:
研究分析羊种布鲁氏菌043新疆流行株中编码脂多糖的WbdA基因和WbkE基因的原核表达以及蛋白的生物信息学信息。以043菌株为模板，构建重组质粒pET-30a-WbdA和pET-30a-WbkE，转化至E.coli BL21(DE3)菌株，IPTG诱导表达，然后通过Western Blot分析检测目的蛋白的表达，最后运用DNAMAN和TMHMM Server v.2.0软件等对WbdA基因和WbkE基因编码的氨基酸序列进行分析。结果显示，成功构建了WbdA基因和WbkE基因的原核表达载体pET-30a-WbdA和pET-30a-WbkE，并且在大肠杆菌中成功表达。生物信息学分析结果显示，043菌株的WbdA蛋白和WbkE蛋白与标准菌株16M的WbdA蛋白和WbkE蛋白的同源性高。WbdA蛋白有1个跨膜结构区，没有信号肽，20个抗原决定簇，二级结构主要由α-螺旋构成，利用Phyre2在线软件成功构建了该蛋白的3D模型。WbkE蛋白没有跨膜结构区，没有信号肽，有17个抗原决定簇，二级结构主要由α-螺旋构成，并利用Phyre2在线软件成功构建了该蛋白的3D模型。
Abstract:
To prokaryotically express WbdA gene and WbkE gene encoding lipopolysaccharide (LPS) from 043 Xinjiang strains of Brucella melitensis, the recombinant plasmids of pET-30a-WbdA and pET-30a-WbkE were constructed with 043 strains as the template and transformed into Escherichia coli BL 21(DE3). The expressions of WbdA protein and WbkE protein were induced by IPTG and detected by Western blotting. Bioinformatics analysis showed that the WbdA protein and WbkE protein of strain 043 shared high homologies with those of 16M strain. WbdA protein has a transmembrane structure and 20 antigenic determinants, but no signal peptide. The secondary structure is consisted of α-helix. Similarly, the WbkE protein has 17 antigenic determinants, but no transmembrane domain and signal peptide. The secondary structure is dominated by α-helix. The 3D models of the two proteins were successfully constructed using Phyre2 online software.

参考文献/References:

［1］FRANCO M P, MULDER M, GILMAN R H, et al. Human brucellosis[J]. The Lancet Infectious Diseases, 2007, 7(12): 775-786.
［2]李军，李常挺，潘艳，等．羊种布鲁氏菌NN1202和LA1105株OPM25和OPM31蛋白B细胞抗原表位分析［J］．江苏农业科学，2015,43(8):38-41.
［3]PAPPAS G, PAPADIMITRIOU P, AKRITIDIS N，et al. The new global map of human brucellosis[J]. The Lancet Infectious Diseases, 2006, 6(2): 91-99.
［4]FICHT T A. Intracellular survival of Brucella: defining the link with persistence[J]. Veterina Microbiol, 2003, 92(3): 213-223.
［5]ELZER P H, HAGIUS S D, DAVIS D S, et al. Characterization of the caprine model for ruminant brucellosis[J]. Veterinary Microbiology, 2002, 90(1): 425-431.
［6]HAMDY M E R, EL-GIBALY S M, MONTASSER A M. Comparison between immune responses and resistance induced in BALB/c mice vaccinated with RB51 and Rev. 1 vaccines and challenged with Brucella melitensis bv. 3[J]. Veterinary Microbiology, 2002, 88(1): 85-94.
［7]GODFROID J, CLOECKAERT A, LIAUTARD J P, et al. From the discovery of the Malta fever’s agent to the discovery of a marine mammal reservoir, brucellosis has continuously been a re-emerging zoonosis[J]. Veterinary Research, 2005, 36(3): 313-326.
［8]胡 森, 步志高. 布氏杆菌病概况及其研究进展[J]. 畜牧兽医科技信息, 2003, 12(9): 63-65.
［9]HAAG A F, MYKA K K, ARNOLD M F F, et al. Importance of lipopolysaccharide and cyclic β-1, 2-glucans in Brucella-mammalian infections[J]. International Journal of Microbiology, 2010, 2010：124509.
［10]HE Y, XIANG Z. Bioinformatics analysis of Brucella vaccines and vaccine targets using VIOLIN[J]. Immunome Research, 2010, 6(1): S5.
［11]MORENO E, CLOECKAERT A, MORIYóN I. Brucella evolution and taxonomy[J]. Veterinary Microbiology, 2002, 90(1): 209-227.
［12]VON BARGEN K, GORVEL J P, SALCEDO S P. Internal affairs: investigating the Brucella intracellular lifestyle[J]. FEMS Microbiology Reviews, 2012, 36(3): 533-562.

相似文献/References:

[1]徐龙,曹小安,刘永生,等.布鲁氏菌BP26蛋白对鼠源树突状细胞分化及抗原提呈作用[J].江苏农业学报,2020,(04):984.[doi:doi:10.3969/j.issn.1000-4440.2020.04.025]
　XU Long,CAO Xiao-an,LIU Yong-sheng,et al.Antigen presentation effect and differentiation of murine dendritic cells stimulated by Brucella BP26 protein[J].,2020,(02):984.[doi:doi:10.3969/j.issn.1000-4440.2020.04.025]

备注/Memo

备注/Memo:
收稿日期：2016-11-24 基金项目：国家“973”计划项目（2010CB530200） 作者简介：王红红(1989-)，女，甘肃天水人，硕士研究生，主要从事动物基因工程研究。（E-mail）1760913825@qq.com 通讯作者：陈创夫，（E-mail）ccf-xb@163.com
更新日期/Last Update: 2017-05-02