Pajunen M, Kiljunen S, Skurnik M
Department of Medical Biochemistry and Molecular Biology, Institute of Biomedicine, University of Turku, Finland.
J Bacteriol. 2000 Sep;182(18):5114-20. doi: 10.1128/JB.182.18.5114-5120.2000.
Bacteriophage phiYeO3-12 is a lytic phage of Yersinia enterocolitica serotype O:3. The phage receptor is the lipopolysaccharide O chain of this serotype that consists of the rare sugar 6-deoxy-L-altropyranose. A one-step growth curve of phiYeO3-12 revealed eclipse and latent periods of 15 and 25 min, respectively, with a burst size of about 120 PFU per infected cell. In electron microscopy phiYeO3-12 virions showed pentagonal outlines, indicating their icosahedral nature. The phage capsid was shown to be composed of at least 10 structural proteins, of which a protein of 43 kDa was predominant. N-terminal sequences of three structural proteins were determined, two of them showing strong homology to structural proteins of coliphages T3 and T7. The phage genome was found to consist of a double-stranded DNA molecule of 40 kb without cohesive ends. A physical map of the phage DNA was constructed using five restriction enzymes. The phage infection could be effectively neutralized using serum from a rabbit immunized with whole phiYeO3-12 particles. The antiserum also neutralized T3 infection, although not as efficiently as that of phiYeO3-12. phiYeO3-12 was found to share, in addition to the N-terminal sequence homology, several common features with T3, including morphology and nonsubjectibility to F exclusion. The evidence conclusively indicated that phiYeO3-12 is the first close relative of phage T3 to be described.
噬菌体phiYeO3-12是小肠结肠炎耶尔森氏菌O:3血清型的一种裂解性噬菌体。该噬菌体的受体是这种血清型的脂多糖O链,其由稀有糖6-脱氧-L-阿洛吡喃糖组成。phiYeO3-12的一步生长曲线显示,其隐蔽期和潜伏期分别为15分钟和25分钟,每个受感染细胞的爆发量约为120个噬菌斑形成单位。在电子显微镜下,phiYeO3-12病毒粒子呈现五边形轮廓,表明其二十面体性质。噬菌体衣壳由至少10种结构蛋白组成,其中一种43 kDa的蛋白占主导。测定了三种结构蛋白的N端序列,其中两种与大肠杆菌噬菌体T3和T7的结构蛋白具有高度同源性。发现噬菌体基因组由一个40 kb的双链DNA分子组成,没有粘性末端。使用五种限制酶构建了噬菌体DNA的物理图谱。用全phiYeO3-12颗粒免疫的兔血清可有效中和噬菌体感染。该抗血清也能中和T3感染,尽管效率不如phiYeO3-12。发现phiYeO3-12除了具有N端序列同源性外,还与T3有几个共同特征,包括形态和对F排斥不敏感。证据确凿地表明,phiYeO3-12是被描述的第一种与噬菌体T3亲缘关系密切的噬菌体。