一株裂解性奶牛乳房炎源屎腸球菌噬菌體的分離及其生物學(xué)特性分析
發(fā)布時間：2019-02-13 13:39
【摘要】：從新疆石河子地區(qū)奶牛糞樣中分離裂解性屎腸球菌噬菌體,對其進(jìn)行純化及生物學(xué)特性分析,為治療和控制細(xì)菌性感染提供基礎(chǔ)。利用雙層平板培養(yǎng)法從奶牛糞樣中分離、純化噬菌體,并觀察噬菌斑特征,將純化后的噬菌體濃縮液用磷鎢酸負(fù)染,通過透射電子顯微鏡觀察其形態(tài)特征。對該噬菌體進(jìn)行全基因組測序和遺傳進(jìn)化分析,用雙層平板培養(yǎng)法測定其宿主譜、最佳感染復(fù)數(shù)、一步生長曲線、熱穩(wěn)定性及pH穩(wěn)定性。結(jié)果如下:分離并純化出一株裂解性屎腸球菌噬菌體,命名為vB_EfaM_XJ3,其噬菌斑圓形透明、無暈環(huán)且邊緣整齊,直徑1.2~1.5mm;電鏡觀察其頭部呈廿面體,頭長約53.15nm,尾長約14.42nm×199.62nm;核酸類型為dsDNA,基因組全長為157.189kb,GC含量為50.27%;其基因序列與沙門菌噬菌體SKML-39的相似性高達(dá)98%。生物學(xué)特性研究顯示vB_EfaM_XJ3目前只裂解其宿主菌;最佳感染復(fù)數(shù)為0.001,潛伏期為15min,爆發(fā)期為175min,裂解量約為84;能耐受50℃左右高溫,在pH 5.0~11.0范圍內(nèi)效價穩(wěn)定。分離鑒定出一株裂解性肌尾科屎腸球菌噬菌體,對它進(jìn)行了生物學(xué)特性和親緣性分析,為沙門菌和屎腸球菌抑菌劑的研究與應(yīng)用奠定了基礎(chǔ)。
[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，
本文編號：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，
本文編號：2421602
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