幽门螺杆菌中国临床分离株iceA基因突变株的构建和鉴定  

Construction and identifi cation of iceA gene mutant strain of Helicobacter pylori isolated from Chinese patients

作　　者：马一君[1,2] 段广才[1,3] 张荣光[1] 张卫东[1] 范清堂[3] 

机构地区：[1]郑州大学公共卫生学院流行病学教研室,河南省郑州市450001 [2]河南中医学院基础医学院,河南省郑州市450008 [3]河南省分子医学重点学科开放实验室,河南省郑州市450052

出　　处：《世界华人消化杂志》2009年第11期1098-1102,共5页World Chinese Journal of Digestology

摘　　要：目的:构建幽门螺杆菌(Hpylori)iceA基因突变株.方法:克隆iceA基因及两侧的部分序列至载体pBluescriptSKⅡ(-)多克隆位点中.运用基因重组方法将卡那霉素抗性基因(kam)插人iceA基因片段之间,构建出带卡那霉素抗性标志的突变载体pBS-iceA-kam.再将突变载体转化至Hpylori中国临床分离株MEL-Hpylori27中,用卡那霉素筛选iceA基因的突变株,经PCR和DNA测序鉴定.结果:用PCR方法扩增突变株基因组DNA,结果显示卡那霉素抗性基因已插入iceA基因片段中,测序进一步证实已筛选获得Hpylori中国临床分离株iceA基因的突变株.结论:成功构建一株HpyloriiceA基因突变株.AIM： To construct an iceA gene mutant strain of Chinese H pylori.METHODS： The iceA gene and its partial upper and downstream flank DNA fragments were cloned into plasmid pBluescript SK II （-）. And kanamycin resistance gene from pEGFP-N2 was inserted into iceA gene to construct objective plasmid pBS-iceA-kam. Then pBS-iceA-kam was transformed into H pylori cells by electroporation and the strains expressing kanamycin resistance genes were selected by kanamycin agar, which were identified by PCR and sequencing analysis. RESULTS： The result of PCR amplified and sequencing analysis from the genome DNA of the constructed mutant strain showed that kanamycin resistance genes were inserted into iceA gene successfully.CONCLUSION： The iceA mutant strain of H pylori isolated from China is constructed successfully.

关 键 词：幽门螺杆菌 ICEA基因 突变株 

分 类 号：R573[医药卫生—消化系统]