产志贺样毒素的噬菌体φ297在大肠杆菌K514染色体上插入位点的定位研究  

Identification of escherichia coli K514 chromosomal insertion sites of Shiga toxin-producing phage φ297

作　　者：翟静[1] 张媛英[1] 王涛[1] 柏素云[1] 孙凌云[1] 蒋汉明[1] 

机构地区：[1]泰山医学院生物化学教研室,山东泰安271000

出　　处：《泰山医学院学报》2005年第1期8-12,共5页Journal of Taishan Medical College

摘　　要：目的　鉴定噬菌体φ2 97在大肠杆菌K5 1 4染色体上的插入位点。方法　采用加接头PCR、分子克隆、亚克隆等分子生物学方法从与噬菌体933W的整合酶基因int同源的核苷酸序列开始向未知区域进行步移测序,寻找目的基因。将获得的DNA序列与核苷酸数据库进行比较。结果　确定了细菌性噬菌体φ2 97的attL、attR和中心序列位于大肠杆菌K5 1 4的yecE 基因内。结论　噬菌体φ2 97在宿主细胞E coliK5 1 4染色体上的整合位点是yecE基因。Objective: To identify the insertion site of prophage φ297 in the host of E. coli K514 chromosome. Methods: Molecular techniques, such as Siebert PCR were used, for walking from the int gene of prophage φ297, which was similar to that of phage 933W to an unknown region in genomic DNA. A special adaptor was ligated to the ends of DNA fragments generated by digestion of genomic DNA with restriction enzymes that generated blunt ended fragments. Clone and reclone of PCR products, DNA sequencing and data analysis were used in this study. Results: The attL and attR and the core sequence were determined. The bacterial attachment site of phage φ297 was located in the yecE gene of E. coli K514. Conclusion: The phage φ297 integrate into the yecE gene of the E. coli K514 genome.

关 键 词：噬菌体φ297 埃希氏大肠杆菌K12 Shiga毒素 附加位点 染色体 插入位点 

分 类 号：Q782[生物学—分子生物学]