鸡大肠杆菌ESBLs基因的克隆及序列分析  

Cloning and Sequence Analysis of ESBLs Gene from Escherichia coli in Chicken

作　　者：张晓凤[1] 张春辉[1] 于顺周 

机构地区：[1]郑州牧业工程高等专科学校,河南郑州450011 [2]上海海利生物药品有限公司,上海201403

出　　处：《中国畜牧兽医》2011年第10期86-88,共3页China Animal Husbandry & Veterinary Medicine

基　　金：河南省教育厅2009年度自然科学研究计划项目"致病性大肠杆菌ESBLs耐药质粒的传播与消除"(2009A230009)

摘　　要：分离纯化来自河南不同地区的鸡大肠杆菌菌株,按照NCCLS标准和双纸片协同试验法进行ESBLs基因表型鉴定。然后根据GenBank已发表的序列设计两对引物,选择阳性表型菌株并提取总DNA,进行PCR、克隆、阳性质粒测序,将测定的序列拼接后和参考菌株ESBLs基因序列比较分析。结果显示,15株大肠杆菌菌株有3株菌株含有β-内酰胺酶,为阳性菌株,这3株与2个参考株核苷酸序列同源性为98.9%～99.7%,氨基酸序列同源性为98.1%～99.2%。3株大肠杆菌阳性菌株序列ESBLs基因型均为TEM型,核苷酸有15个位点发生突变,其中6个位点突变引起氨基酸变异。To Purified the isolated E.coli in chickens from different area in Henan,screen PhenotyPe for ESBLs according to NCCLS standards and the double-disc synergy method.Then according to sequences Published in GenBank,designed two Pairs of Primers.The bacteria Producing ESBLs were chose,extracted the total DNA,amPlified with PCR,cloned,sequenced and analyzed.The results showed the three determinated sequences had 98.9% to 99.7% nucleotide sequence homology with the two reference sequence,amino homology was 98.1% to 99.2%.And there were 15 nucleotide mutations and 6 mutations of amino in the 3 determinated sequences.

关 键 词：ESBLs基因 克隆 序列分析 

分 类 号：S852.61[农业科学—基础兽医学]