摘要
为构建中国幽门螺杆菌 (Hp)致病岛缺失突变的载体 ,采用PCR扩增、质粒抽提、琼脂糖凝胶电泳、限制性酶切、目的基因与载体的连接、感受态细胞的制备及转化等多种基因工程技术 ,将氯霉素抗性基因CmR 连接到PCR扩增致病岛两端区域产生的两目的基因片段之间 ,并共同插入到pBluescriptSK 载体的多克隆位点中 ,经氨苄青霉素抗性筛选及相应的限制性酶切等方法鉴定。构建的突变载体经内切酶酶切分析显示 :产生的条带与设计结果完全一致。
A vector for a mutant Chinese Helicobacter pylori (Hp) strains knock out pathogenicity island (PAI) was constructed, which was the basis to establish a Chinese Hp mutant deleting pathogenicity island. Genetic engineering techniques such as polymerase chain reaction, plasmid extraction, agarose gel electrophoresis, restriction analysis, ligation, preparation of competence cell and transformation were used to make two cloning fragments containing two end regions of PAI and a selectable chloramphenicol resistance marker between them, and then engineering the recombined fragments into pBluescript plasmid. The result showed that restriction analysis demonstrated that the engineering mutant vector had been recombined. It suggested that we constructed an engineering mutant vector which targeted to delete the PAI in Chinese Hp strains. It will be useful for addressing the role of PAI in the pathogenesis of Hp infection.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2001年第11期798-800,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金资助课题 (编号 396 70 6 48)
欧共体委员会科学基金资助课题 (编号IC18CT95 0 0 2 4)
