Johnsen M G, Neve H, Vogensen F K, Hammer K
Danish Center for Lactic Acid Bacteria, Department of Dairy and Food Science, Royal Veterinary and Agricultural University, Frederiksberg C.
Virology. 1995 Oct 1;212(2):595-606. doi: 10.1006/viro.1995.1517.
The major proteins of phage TP901-1 virion were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and structural relations were determined using specific antibodies, obtained by affinity purification from a polyclonal serum. A 23-kDa protein was identified as the major tail protein, and a 31-kDa molecule as the major head protein, respectively. Labeling experiments with antibodies against two proteins, with molecular masses of 20 and 19 kDa, indicated that they were baseplate-related components. A 72-kDa protein was found to be part of a neck passage structure, which includes a collar. Evidence for the presence of attached whiskers was also obtained. T7 RNA polymerase-mediated expression of the two major proteins confirmed the gene location of the previously sequenced region of the phage genome. The relation to other lactococcal phages was determined by DNA hybridization and antibody probing, showing that despite low DNA similarity, TP901-1 NPS epitopes were detected in both related and unrelated small isometric-headed phages.
通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳对噬菌体TP901-1病毒粒子的主要蛋白质进行了表征,并使用从多克隆血清中通过亲和纯化获得的特异性抗体确定了其结构关系。分别鉴定出一种23 kDa的蛋白质为主要尾蛋白,一种31 kDa的分子为主要头部蛋白。用针对两种分子量分别为20 kDa和19 kDa的蛋白质的抗体进行标记实验,表明它们是与基板相关的成分。发现一种72 kDa的蛋白质是颈部通道结构的一部分,该结构包括一个衣领。还获得了附着须状物存在的证据。T7 RNA聚合酶介导的两种主要蛋白质的表达证实了噬菌体基因组先前测序区域的基因定位。通过DNA杂交和抗体探测确定了与其他乳球菌噬菌体的关系,结果表明,尽管DNA相似性较低,但在相关和不相关的小等轴头噬菌体中均检测到了TP901-1 NPS表位。