不同动物源性大肠杆菌多重耐药调控基因rob的同源性分析  

Homology analysis of multidrug-resistance regulating gene rob of Escherichia coli from different animal species

作　　者：马红霞[1] 刘玉堂[1] 伞治豪[1] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《中国兽医科学》2007年第12期1035-1040,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30400326);吉林省杰出青年基金项目(20050124);吉林省科技厅项目(20030425-03)

摘　　要：为研究大肠杆菌多重耐药调控基因rob与不同动物源性及多重耐药水平之间的关系,选取临床分离的不同动物源性大肠杆菌5株及大肠杆菌药敏质控株ATCC25922,在对其进行主要治疗药物耐药性检测的基础上,分别以其染色体DNA为模板,通过PCR反应扩增出大肠杆菌多重耐药调控基因rob,将该基因分别与克隆载体pMD18-T连接,连接产物转化至大肠杆菌JM109感受态细胞中,对经酶切与PCR反应鉴定为阳性的克隆质粒进行了核苷酸序列测定。测序结果表明:不同动物源性大肠杆菌的rob基因与Gen-Bank中该基因的核苷酸序列及所推导的氨基酸序列的同源性较高。不同源性大肠杆菌的rob基因的核苷酸序列与其动物源性有关,该基因的核苷酸序列的个别突变位点可能影响AcrAB外输泵的表达水平,进而影响其多重耐药水平。In order to study the relationship between multidrug-resistance regulating genes rob of Escherichia coli from different animal species and levels of multidrug-resistance,5 strains of E. coli isolated from different animal species and control strain ATCC25922 were selected. Based upon drug-resistance detection of main therapeutic drugs,rob genes of multidrug-resistance of E. coli were amplified by PCR from chromosome DNA of E. coli and ligated into pMD18-T Simple Vector. The ligated products were translated into E. coli JM109. A positive plasmid identified by enzymatic digestion and PCR was sequenced. The sequencing result indicated that homology of nucleotide sequences of and deduced amino acids from the rob genes between the 5 E. coli isolates and that of GenBank were high. Some gene mutations of rob gene of the 5 E. coli isolates were related to animal species and to the level of drug resistance by affecting on the expression level of AcrAB efflux pump.

关 键 词：大肠杆菌 多重耐药 rob基因 克隆 同源性分析 

分 类 号：S852.612[农业科学—基础兽医学] Q523.8[农业科学—兽医学]