一株撒坝猪源大肠杆菌噬菌体的生物学特性研究  

Study on The Biological Characteristics of an E.coli Bacteriophage from Saba pigs

作　　者：万全 张博 高飞龙 单春兰 赵维薇 邓静 王喜 赵汝 高利波 高洪 WAN Quan;ZHANG Bo;GAO Feilong;SHAN Chunlan;ZHAO Weiwei;DENG Jing;WANG Xi;ZHAO Ru;GAO Libo;GAO Hong(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China;College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;College of Food Science and Technology,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学动物医学院,云南昆明650201 [2]云南农业大学动物科学技术学院,云南昆明650201 [3]云南农业大学食品科学技术学院,云南昆明650201

出　　处：《家畜生态学报》2021年第7期47-51,共5页Journal of Domestic Animal Ecology

基　　金：国家自然科学基金资助项目(31260594);云南省现代农业生猪产业技术体系建设(云财农[2009]171号);云南省高校科技创新团队支持计划资助(云教科[2011]14号)

摘　　要：为了解撒坝猪源大肠杆菌噬菌体分离株的生物学特性。自撒坝猪源大肠杆菌中分离到一株裂解性噬菌体,利用PCR扩增噬菌体衣壳蛋白gp23基因,并通过同源性比对及遗传进化分析来鉴定噬菌体。经增殖纯化培养后测定噬菌体的效价和对菌株的裂解效应,而后测定其最佳感染复数、热稳定性、pH稳定性及绘制生长曲线以探究该噬菌体的生物学特性。结果表明,分离株衣壳蛋白gp23基因与噬菌体slur14亲缘关系最近,属于有尾噬菌体目、肌尾噬菌体科T4-like噬菌体;分离株具有较高的效价,达8.67×10^(9)pfu/m;针对实验室保存的92株已确定血清型的大肠杆菌裂解率为61.95%,完全裂解率为27.17%;在60℃条件下作用30 min,或在pH<5和pH>9时,噬菌体的效价表现明显下降;生长曲线绘制后可知该噬菌体感染宿主菌的潜伏时间约为30 min,爆发时间约为120 min,裂解量为41 pfu/cell。该试验成功分离得到裂解能力较强、裂解谱较广且对理化因素耐受力较强的T4-like噬菌体。To understand the biological characteristics of E.coli bacteriophage isolated from Saba pigs,a lysogenic bacteriophage derived from Saba pigs was isolated from E.coli.The phage capsid protein gp 23 gene was amplified by PCR and identified by homology comparison and genetic evolution analysis.After proliferation,purification and cultivation,the bacteriophage titer and the lysis effect on the strain were measured,and then the optimal infection complex number,thermal stability,pH stability and growth curve were drawn to explore the biological characteristics of the bacteriophage.The results showed that the gp23 gene of the capsid protein of the isolate had the closest genetic relationship with the bacteriophage slur14,which belonged to the T4-like bacteriophage of the order of tail bacteriophage and myobacteriophage.The isolate had a high titer,reaching 8.67×10^(9) pfu/m.For the 92 strains of E.coli with determined serotypes stored in the laboratory,the lysis rate was 61.95%and the complete lysis rate was 27.17%.The bacteriophage's titer was obviously reduced,when the pH<5 and pH>9 or at 60℃for 30 min.From the growth curve,we could find that the incubation time of the phage to infect the host bacteria was about 30 min,the outbreak time was about 120 min,and the lysis amount was 41 pfu/cell.In this experiment,a T4-like bacteriophage with strong lysis ability,broad lysis spectrum and strong tolerance to physical and chemical factors was successfully isolated.

关 键 词：大肠杆菌 噬菌体 分离鉴定 生物学特性 

分 类 号：Q939.48[生物学—微生物学]