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目的分离广谱大肠杆菌噬菌体并对其进行种属的鉴定。方法以不同种临床致病性大肠杆菌为指示菌,从未经处理的污水样品中分离噬菌体;采用蛋白酶K/SDS的方法提取噬菌体基因组,制备重组载体,将阳性重组质粒进行测序;将测序结果进行BLAST,推测该噬菌体种属分类位置。结果分别以大肠杆菌E1~E17共17种细菌作为指示菌,成功分离出一种广谱噬菌体,可裂解31株临床分离致病性大肠杆菌中的13株,并将其命名为IME11;由噬菌体基因组限制性酶切片段分析表明其遗传物质为dsDNA;将噬菌体IME11基因组的2个随机片段测序结果进行BLAST,推测其属于N4-like噬菌体属。结论分离出了一种广谱噬菌体,在分类学上属于N4-like噬菌体属。
Objective To isolate a broad spectrum of E. coli phage and identify its species. Methods Phage were isolated from untreated sewage samples by using different kinds of clinical pathogenic E.coli as indicator bacteria. The phage genome was extracted by protease K / SDS, and the recombinant vector was prepared. The positive recombinant plasmids were sequenced. BLAST, speculated that the phage taxonomic location. Results A total of 17 kinds of bacteria from E. coli E1 ~ E17 were used as the indicator bacteria, respectively. A broad-spectrum bacteriophage was successfully isolated and 13 strains isolated from 31 clinical isolates of pathogenic E. coli were isolated and named as IME11. The restriction fragment analysis of the genome showed that the genetic material was dsDNA. Two random fragments of the phage IME11 genome were BLASTed and sequenced to belong to the N4-like phage genus. Conclusion A broad-spectrum bacteriophage was isolated and taxonomically belongs to the N4-like bacteriophage genus.