单增李斯特菌溶源性噬菌体的诱导及鉴定

Induction and Identification of Listeria monocytogenes Lysogenic Phage

利用丝裂霉素C诱导获得单增李斯特菌溶源性噬菌体,并确定其核酸类型。PCR及其扩增产物的序列测定检测原噬菌体,透射电子显微镜观察丝裂霉素C诱导获得的溶源性噬菌体颗粒,DNase I、RNase A和绿豆核酸酶分别消化噬菌体基因组,确定噬菌体的核酸类型。结果显示,38株单增李斯特菌分离株中的16株在tRNAArg位点携有原噬菌体,其中1株可诱导出噬菌体颗粒,该噬菌体为长尾噬菌体,核酸为dsDNA。噬菌体的成功诱导为噬菌体及其功能基因的开发利用奠定了基础。

To induce lysogenic phage from Listeria monocytogenes with Mitomycin C,and determine nucleic acid type of the induced phage,detection of prophages at tRNAArg attach sites of bacterial chromosomes was carried out by Polymerase Chain Reaction amplification and subsequent sequencing the PCR products,phage particles were observed by transmission electronic microscope,phage nucleic acid was digested with nucleases.The results showed that 16 of 38 Listeria monocytogenes isolates carried pro-phages at tRNAArg attachment sites,a lysogenic phage with typical shape of Siphovirus,named W001,was successfully induced from one of the above 16 Listeria monocytogenes isolates.After digestion of the extracted phage nucleic acid with DNase I,RNase A and Mung Bean nuclease,respectively,the nucleic acid type of phage W001 was determined to be double-stranded DNA （dsDNA）.The sucessful induction of the phage laid foundations for the devolpement of phage and its functional genes.