大肠杆菌外膜蛋白ompF基因与Ⅰ类整合酶基因在生物被膜状态下表达相关性的研究  

Correlation between ompF gene of Escherichia coli outer membrane protein and expression of class Ⅰ integrase gene in biofilm state

作　　者：王艺凝 周建华[1] 马丽娜[1] 李学瑞[1] 刘永生[1] WANG Yi-ning;ZHOU Jian-hua;MA Li-na;LI Xue-rui;LIU Yong-sheng(Key Laboratory of Grazing Animal Disease of Ministry of Agriculture/State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室/农业部草食动物疫病重点开放实验室

出　　处：《中国兽医学报》2020年第1期109-115,161,共8页Chinese Journal of Veterinary Science

基　　金：甘肃省科技计划资助项目(17YF1WA170)

摘　　要：本试验拟研究生物被膜状态下外膜蛋白ompF基因与大肠杆菌Ⅰ类整合酶基因表达的相关性,初步探讨外膜蛋白对Ⅰ类整合酶基因的调控作用。利用Red同源重组系统构建大肠杆菌野生株的ompF基因缺失株和回复株,分别检测它们的生长曲线,并通过实时荧光定量PCR(RT-PCR)检测生物被膜状态下亲本株,ompF基因缺失株和回复株的Ⅰ类整合酶基因mRNA的表达水平,分析它们的表达相关性。生长曲线检测发现ompF基因虽然不是大肠杆菌生长繁殖所必需的功能性基因,敲除后不会导致大肠杆菌的死亡,但会对大肠杆菌的生长有一定的影响。生物被膜状态下ompF基因缺失株Ⅰ类整合酶基因的表达水平比野生株明显降低,回复株Ⅰ类整合酶基因的表达水平有所升高,但未回复到野生株的表达水平。初步确定在生物被膜状态下,ompF基因对大肠杆菌Ⅰ类整合酶基因的表达有影响,此结果可为进一步研究生物被膜状态下大肠杆菌的耐药机制开拓新的研究方向。This study was to investigate the correlation between the outer membrane protein ompF gene and the expression of Escherichia coli(E.coli) class I integrase gene in biofilm state.The Red homologous recombination system was used to construct the ompF mutuant strain and the complemented strain,detected their growth curves separately.Then the real-time fluorescence quantitative PCR(RT-PCR) was used to detect the expression level of the integrase gene mRNA of wild type,ompF mutuant and restore strain in biofilm state and analyzed their expression correlation.The growth curve test found that ompF gene is not a necessarily functional gene for the growth and reproduction of E.coli,and does not cause death of E.coli after knockout,but has a certain influence on the growth of E.coli.The expression level of class I integrase gene of ompF mutuant strain was significantly lower than that of wild-type strain under the condition of biofilm,and the expression level of class I integrase gene of restore strain was increased,but did not revert to the previous expression level.It was initially determined that the ompF gene has an effect on the expression of E.coli class Ⅰ integrase gene in the biofilm state,this result can open up a new research direction for further study of the resistance mechanism of E.coli in biofilm state.

关 键 词：大肠杆菌 ompF基因 Ⅰ类整合酶 耐药基因 

分 类 号：S855.12[农业科学—临床兽医学]