一株裂解性奶牛乳房炎源屎肠球菌噬菌体的分离及其生物学特性分析
发布时间：2019-02-13 13:39
【摘要】：从新疆石河子地区奶牛粪样中分离裂解性屎肠球菌噬菌体,对其进行纯化及生物学特性分析,为治疗和控制细菌性感染提供基础。利用双层平板培养法从奶牛粪样中分离、纯化噬菌体,并观察噬菌斑特征,将纯化后的噬菌体浓缩液用磷钨酸负染,通过透射电子显微镜观察其形态特征。对该噬菌体进行全基因组测序和遗传进化分析,用双层平板培养法测定其宿主谱、最佳感染复数、一步生长曲线、热稳定性及pH稳定性。结果如下:分离并纯化出一株裂解性屎肠球菌噬菌体,命名为vB_EfaM_XJ3,其噬菌斑圆形透明、无晕环且边缘整齐,直径1.2~1.5mm;电镜观察其头部呈廿面体,头长约53.15nm,尾长约14.42nm×199.62nm;核酸类型为dsDNA,基因组全长为157.189kb,GC含量为50.27%;其基因序列与沙门菌噬菌体SKML-39的相似性高达98%。生物学特性研究显示vB_EfaM_XJ3目前只裂解其宿主菌;最佳感染复数为0.001,潜伏期为15min,爆发期为175min,裂解量约为84;能耐受50℃左右高温,在pH 5.0~11.0范围内效价稳定。分离鉴定出一株裂解性肌尾科屎肠球菌噬菌体,对它进行了生物学特性和亲缘性分析,为沙门菌和屎肠球菌抑菌剂的研究与应用奠定了基础。
[Abstract]:The bacteriophage of Enterococcus faecium was isolated from cow dung in Shihezi area of Xinjiang. The bacteriophage was purified and its biological characteristics were analyzed so as to provide a basis for the treatment and control of bacterial infection. The phage was purified from cow dung by double layer plate culture, and the plaque characteristics were observed. The purified concentrated phage was negatively stained with phosphotungstic acid, and its morphological characteristics were observed by transmission electron microscope (TEM). The whole genome of the phage was sequenced and the genetic evolution was analyzed. The host spectrum of the phage was determined by double-layer plate culture method. The best complex number of infection, one-step growth curve, thermal stability and pH stability were obtained. The results were as follows: a lytic Enterococcus faecium bacteriophage was isolated and purified. It was named vB_EfaM_XJ3, with circular and transparent plaque, no halo ring and neat edges, with a diameter of 1.2 ~ 1.5 mm. The head length was about 53.15nm, the tail length was about 14.42nm 脳 199.62nm.The nucleic acid type was dsDNA, genome with a total length of 157.189kb ~ (1) and GC content of 50.27nm, and the length of its head was 53.15nm, and the length of its tail was about 14.42nm 脳 199.62nm. The similarity between its gene sequence and Salmonella phage SKML-39 is as high as 98%. The biological characteristics showed that vB_EfaM_XJ3 only lyses its host bacteria at present, the optimum number of infection is 0.001, the incubation period is 15 min, the outbreak period is 175 min, the amount of cleavage is about 84; It can withstand about 50 鈩，
本文编号：2421602
[Abstract]:The bacteriophage of Enterococcus faecium was isolated from cow dung in Shihezi area of Xinjiang. The bacteriophage was purified and its biological characteristics were analyzed so as to provide a basis for the treatment and control of bacterial infection. The phage was purified from cow dung by double layer plate culture, and the plaque characteristics were observed. The purified concentrated phage was negatively stained with phosphotungstic acid, and its morphological characteristics were observed by transmission electron microscope (TEM). The whole genome of the phage was sequenced and the genetic evolution was analyzed. The host spectrum of the phage was determined by double-layer plate culture method. The best complex number of infection, one-step growth curve, thermal stability and pH stability were obtained. The results were as follows: a lytic Enterococcus faecium bacteriophage was isolated and purified. It was named vB_EfaM_XJ3, with circular and transparent plaque, no halo ring and neat edges, with a diameter of 1.2 ~ 1.5 mm. The head length was about 53.15nm, the tail length was about 14.42nm 脳 199.62nm.The nucleic acid type was dsDNA, genome with a total length of 157.189kb ~ (1) and GC content of 50.27nm, and the length of its head was 53.15nm, and the length of its tail was about 14.42nm 脳 199.62nm. The similarity between its gene sequence and Salmonella phage SKML-39 is as high as 98%. The biological characteristics showed that vB_EfaM_XJ3 only lyses its host bacteria at present, the optimum number of infection is 0.001, the incubation period is 15 min, the outbreak period is 175 min, the amount of cleavage is about 84; It can withstand about 50 鈩，
本文编号：2421602
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