作 者：
丁雪燕;张琪;田延;王亨;张伟;石宝兰;张建军;朱国强
单 位：
江苏省动物重要疫病与人兽共患病防控协同创新中心;扬州大学兽医学院;国药集团扬州威克生物工程有限公司
关键词：
Red同源重组系统;Ⅵ型分泌系统;hcp1;hcp2
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/phcp1phcp2。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。
译 名：
Construction of the type Ⅵ secretion system genes of hcp1 and hcp2 deleted strains from the avian pathogenic Escherichia coli CE129
作 者：
DING Xue-yan;ZHANG Qi;TIAN Yan;WANG Heng;ZHANG Wei;SHI Bao-lan;ZHANG Jian-jun;ZHU Guo-qiang;College of Veterinary Medicine, Yangzhou University;Jiangsu Co-Innovation Center for Important Animal Infectious Diseases and Zoonoses;Yangzhou Vacbio Bioengineering Co., Ltd;
关键词：
Red recombination system;;type Ⅵ secretion system;;hcp1;;hcp2
摘 要：
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 p BR322 or p ACYC184 to construct the complementary strains of CE129 △hcp1/phcp1, CE129 △hcp2/phcp2 and CE129 △hcp1 △hcp2/phcp1 phcp2,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.
