多株幽门螺杆菌肽脱甲酰基酶基因克隆及序列比较  

Cloning and sequence alignment of peptide deformylases from various strains of Helicobacter pylori

作　　者：幺山山[1] 曾明[1] 王斌[1] 段海清[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《中华微生物学和免疫学杂志》2005年第3期176-178,共3页Chinese Journal of Microbiology and Immunology

摘　　要：目的　为开发以幽门螺杆菌 (Hp)肽脱甲酰基酶为靶位的新药 ,首先考察该基因的保守性。方法　从我国不同地区分离、选择多株Hp ,培养并提取细菌DNA ,设计特异引物以PCR方法扩增其肽脱甲酰基酶基因 (def) ,并克隆到 pGEMT Easy载体中 ,转化大肠杆菌 ,提取质粒鉴定重组质粒并测序。结果　经序列比较 ,发现def基因有高度保守性 ,在重要的基序点未发生变异。结论　本研究为以肽脱甲酰基酶作为靶位筛选药物提供了依据。Objective To verify gene conservation of peptide deformylases (PDF) from H.pylori . Methods Ten strains from different regions of China were isolated and used to clone this gene. Bacterial DNA was extracted and used as a template to amplify def gene by PCR. def genes obtained were subsequently cloned into pGEM T easy vector and recombinants were identified by enzyme digestion. Results Sequence alignment shows that this gene is extremely conserved in H.pylori . There is no variation in three important motifs related to its function. Conclusion PDF may be an ideal target for new drug development.

关 键 词：肽脱甲酰基酶 序列比较 基因克隆 幽门螺杆菌(Hp) 细菌DNA PCR方法 不同地区 特异引物 大肠杆菌 重组质粒 药物提供 保守性 酶基因 靶位 提取 

分 类 号：R346[医药卫生—基础医学]