鲍曼不动杆菌噬菌体生物学特性的研究  

Biologic characteristics of Bacteriophages specific to Acinetobacter baumannii

作　　者：余静丹[1] 史红艳[1] 王丹[1] 徐花[1] 孙延波[1] 

机构地区：[1]吉林大学白求恩医学院病原生物学系,吉林长春130021

出　　处：《微生物学杂志》2013年第2期17-23,共7页Journal of Microbiology

基　　金：国家自然科学基金(81150037)

摘　　要：从污水中分离鉴定鲍曼不动杆菌噬菌体,并对其生物学特性进行分析,为开发针对细菌感染的噬菌体生物制剂提供前期工作。采用双层琼脂法分离可裂解鲍曼不动杆菌的噬菌体,通过负染法电镜观察噬菌体的大小和形态,将噬菌体和宿主菌以不同比例混合,测定噬菌体的最佳感染复数并观察一步生长曲线,提取噬菌体核酸进行酶切电泳分析,通过SDS-PAGE分析噬菌体的结构蛋白和非结构蛋白。成功分离3株可裂解鲍曼不动杆菌的噬菌体(分别命名为ФAb-1、ФAb-2和ФAb-3),电镜显示噬菌体的头部呈二十面体,直径约50nm,有一短尾。噬菌体ФAb-1的最佳感染复数为10-2,一步生长曲线表明噬菌体在裂解宿主菌时,潜伏期为20 min,爆发期为30 min,裂解量为190 PFU/cell。限制性酶切电泳显示噬菌体ФAb-1的基因组大小约40 kb左右,为双股环状DNA。SDS-PAGE的噬菌体蛋白电泳包括7种蛋白,分子量在29～116 ku。噬菌体ФAb-1、ФAb-2和ФAb-3对鲍曼不动杆菌具有很强的裂解毒性。根据其形态、结构和噬菌体分类法,鲍曼不动杆菌噬菌体属于有尾病毒目,足尾病毒科。Bacteriophages specific to Acinetobacter baumannii （BAB） isolated from sewage was investigated for its bi-ological characteristics to provide early stage＇s work for the development of bio-preparation of phage aiming at bacterial infection. Splittable BAB was isolated and identified using double-layer-agar method. Its size and morphology was ob- served and examined by electron microscopy using negative-staining. The optimal multiplicity of infection and one-step growth curve of the phages were measured by mixing host bacteria and the phages in different proportions. The extrac-ted genomes of the phages were evaluated on electrophoresis and digested with restriction enzymes. SDS-PAGE was employed for analysis of the structural and non-structural proteins of the phages. And it was successfully isolated three splittable BAB respectively named as ФAb-1, ФAb-2 and ФAb-3, transmission electron microscopy showed that head of the phages assumed an icosahedra, 50 nm in diameter, having a short tail. The multiplicity of infection of phage ФAb-1 was 10-2, and one-step growth curve revealed that the latent phase was 20 min, and the eruptive 30 min, and the splitting amount of phage ФAb-1 was 190 PFU/infected cells. Restriction enzyme electrophoresis showed that the size of genome of phage ФAb-1 was about 40 kb with a circular double-stranded DNA. SDS-PAGE analysis indicated protein electrophoresis of the phage included 7 bands with molecular weight around 29 - 116 ku. Phage ФAb-1, qФAb- 2 and ФAb-3 had very strong splitting virulence against A. baumannii, and it was isolated and identified according its morphology, structure, and phage taxonomy. A. baumannii belonged to the order of Caudovirales, and the family of Podoviridae.

关 键 词：鲍曼不动杆菌 噬菌体 一步生长曲线 

分 类 号：Q939.48[生物学—微生物学]