摘要
鉴定大肠杆菌O15 7产生Shiga毒素 (Stx)的噬菌体 φ2 97在大肠杆菌K12染色体上的插入位点。采用加接头PCR方法从与噬菌体 933W的整合酶基因int同源的核苷酸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 ,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K12的yecE基因内。噬菌体 φ2 97在宿主细胞E .coliK12染色体上的整合位点是yecE基因。
The specific integration site of prophage φ297 in the host of E. coli K12 chromosome was identified using molecular techniques. Siebert PCR run 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 generates blunt ended fragments. Clone and reclone of PCR products, DNA sequencing and data analysis were used in this study. Comparison the sequence obtained with the nucleotide database was allowed to determine the attL and attR and the core sequence. The bacterial attachment site of phage φ297 was located in the yecE gene of E. coli K12.The phage φ297 integrates into the yecE gene of the E. coli K12 genome.
出处
《微生物学报》
CAS
CSCD
北大核心
2004年第5期605-608,共4页
Acta Microbiologica Sinica
