BACTERIOPHAGE EPSILON 15

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 噬菌体 Epsilon15 是一种通用转导噬菌体，可感染人/动物病原体肠沙门氏菌 Anatum 血清型。 Epsilon 15 因其识别和结合 O 抗原的能力以及在溶原状态下改变宿主脂多糖的能力而受到研究。负责 O 抗原识别的病毒蛋白是尾刺蛋白。该病毒体包含至少六种结构蛋白和已知序列的大约 40 kb dsDNA 基因组。在序列水平上，结构蛋白与伯克霍尔德杆菌（一种影响囊性纤维化患者的人类病原体）的 Bcep 噬菌体最相似。病毒体结构蛋白和染色体共同形成质量约为 66 兆道尔顿、直径约为 650 的颗粒。乔恩·金的实验室使用质谱方法来识别那些编码病毒颗粒结构蛋白的开放阅读框。这种病毒的结构将揭示结构蛋白的排列，特别是那些更直接参与宿主附着和DNA进入细胞质的亚基的配置。目前正在进行一种遗传方法来分离病毒结构蛋白中的琥珀突变体。这些突变噬菌体感染期间形成的大分子亚组合将通过重建进行纯化和成像。这些突变噬菌体还将用作对病毒与宿主表面的脂多糖和受体蛋白之间的相互作用进行成像的试剂。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacteriophage Epsilon15 is a generalized transducing phage infecting the human/animal pathogen Salmonella enterica serovar Anatum. Epsilon 15 has been studied for its ability to recognize and bind O-antigen as well as its capacity to alter host lipopolysaccharide in the lysogenic state. The viral protein responsible for O-antigen recognition is the tailspike protein. The virion contains at least six structural proteins and an approximately 40 kb dsDNA genome of known sequence. At the sequence level, the structural proteins most closely resemble the Bcep phages of Burkholderia, a human pathogen affecting cystic fibrosis patients. The virion structural proteins and chromosome together make a particle with a mass of approximately 66 Megadaltons and a diameter of roughly 650¿. Jon King's lab has used mass spectrometric methods to identify those open reading frames encoding virion structural proteins. The structure of this virus will reveal the arrangement of structural proteins and, in particular, the configuration of those subunits more directly involved in attachment to the host and passage of DNA into the cytoplasm. A genetic approach is now underway to isolate amber mutants in virus structural proteins. Macromolecular subassemblies formed during infections with these mutant phage will be purified and imaged by reconstruction. These mutant phage will also serve as reagents for imaging the interactions between virus and lipopolysaccharide and receptor proteins at the surface of the host.