[1]丁雪燕?,张 琪?,田 延,等.禽致病性大肠杆菌CE129 VI型分泌系统主要亚单位基因hcp1、hcp2缺失株的构建与验证[J].中国预防兽医学报,2018,(05):381-385.[doi:10.3969/j.issn.1008-0589.201710001]
 DING Xue-yan,ZHANG Qi,TIAN Yan,et al.Construction of the type VI secretion system genes of hcp1 and hcp2 deleted strains from the avian pathogenic Escherichia coli CE129[J].Chinese journal of preventive veterinary medicine,2018,(05):381-385.[doi:10.3969/j.issn.1008-0589.201710001]
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禽致病性大肠杆菌CE129 VI型分泌系统主要亚单位基因hcp1、hcp2缺失株的构建与验证()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年05
页码:
381-385
栏目:
病原生物学
出版日期:
2018-05-15

文章信息/Info

Title:
Construction of the type VI secretion system genes of hcp1 and hcp2 deleted strains from the avian pathogenic Escherichia coli CE129
文章编号:
1008-0589(2018)5-0381-05
作者:
 

丁雪燕12?张 琪12?田 延12王 亨12张 伟123石宝兰123张建军123朱国强12*

 (1. 扬州大学 兽医学院,江苏 扬州 225009;2. 江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏 扬州 225009;
3. 国药集团 扬州威克生物工程有限公司,江苏 扬州 225009)
Author(s):
 

DING Xue-yan12? ZHANG Qi12? TIAN Yan12 WANG Heng12 ZHANG Wei123 SHI Bao-lan123 ZHANG Jian-jun123 ZHU Guo-qiang12*

 (1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; 2. Jiangsu Co-Innovation Center for Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China; 3. Yangzhou Vacbio Bioengineering Co., Ltd, Yangzhou 225009, China)
关键词:
Red同源重组系统VI型分泌系统hcp1hcp2
Keywords:
 Red recombination system  type VI secretion system  hcp1  hcp2
分类号:
S852.6
DOI:
10.3969/j.issn.1008-0589.201710001
文献标志码:
A
摘要:
为获得禽致病性大肠杆菌(APEC) CE129 VI型的hcp1、hcp2基因缺失株,本研究利用Red同源重组系统对APEC野生株CE129的VI型分泌系统(T6SS) hcp1和hcp2基因进行敲除,获得hcp1、hcp2基因单缺失株CE129△hcp1、CE129△hcp2和双缺失株CE129△hcp1△hcp2。将hcp1、hcp2基因分别克隆到表达载体pBR322和pACYC184中,构建相应的基因回补株CE129△hcp1/phcp1、CE129△hcp2/phcp2和CE129△hcp1△hcp22/phcp1 phcp2。通过PCR特异性检测和基因测序显示,上述突变株与回补株均成功构建,且均能够稳定遗传。上述基因缺失株和回补株的成功构建,有助于深入了解APEC CE129菌株T6SS的作用机制,为进一步研究APEC的致病机理及其防控奠定了基础。
  
Abstract:
To construct the deleted hcp1 and hcp2 genes of type VI secretion system (T6SS) of avian pathogenic Escherichia coli (APEC), the CE129 strain of APEC was used as the prototype strain. Then, the single gene deleted strains (CE129△hcp1 and CE129△hcp2) and double gene deleted strain CE129△hcp1△hcp2 from APEC CE129 were prepared using the Red recombination system. In addition, the hcp1 and hcp2 genes were cloned into the expression vectors of pBR322 or pACYC184 to construct the complementary strains of CE129△hcp1/phcp1, CE129△hcp2/phcp2 and CE129△hcp1△hcp2/phcp1phcp2, respectively. These mutant strains and complementary strains were proved to be genetically stable confirmed by PCR and DNA sequencing. This work provided the basis to investigate the mechanism of T6SS in APEC CE129 and understand the pathogenesis of APEC. It might also help in the prevention and control of APEC-caused avian diseases.

参考文献/References:

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                                               (本文编辑:彭永刚)

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更新日期/Last Update: 2018-05-15