论文部分内容阅读
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.
Objective: To construct a mutant of iceA gene of Helicobacter pylori (Hpylori) .Methods: The gene fragment of iceA was cloned into the multiple cloning site of vector pBluescriptSKⅡ (-), and the kanamycin resistance gene ( kam) was inserted into the iceA gene fragment to construct the mutant vector pBS-iceA-kam with kanamycin resistance marker.Mutant vector was transformed into Hpylori Chinese clinical isolate MEL-Hpylori27 and treated with kanamycin The mutant of iceA gene was screened and identified by PCR and DNA sequencing.Results: The genomic DNA of the mutant was amplified by PCR and the results showed that the kanamycin resistance gene was inserted into the iceA gene fragment, and sequencing further confirmed that Hpylori China Clinical isolates of iceA gene mutant.Conclusion: A HpyloriiceA gene mutant was successfully constructed.