Department of Biochemistry and Molecular Biology, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA 19107, USA.
Department of Microbiology, University of Alabama at Birmingham, 845 19th Street South, Birmingham, AL 35294, USA.
STAR Protoc. 2021 Nov 16;2(4):100960. doi: 10.1016/j.xpro.2021.100960. eCollection 2021 Dec 17.
Bacteriophages of the family densely package their genomes into precursor capsids alongside internal virion proteins called ejection proteins. In phage T7 these proteins (gp14, gp15, and gp16) are ejected into the host envelope forming a DNA-ejectosome for genome delivery. Here, we describe the purification and characterization of recombinant gp14, gp15, and gp16. This protocol was used for high-resolution cryo-EM structure analysis of the T7 periplasmic tunnel and can be adapted to study ejection proteins from other phages. For complete details on the use and execution of this protocol, please refer to Swanson et al. (2021).
科噬菌体将其基因组与内部病毒蛋白（称为排出蛋白）一起紧密包装到前衣壳中。在噬菌体 T7 中，这些蛋白质（gp14、gp15 和 gp16）被排出到宿主包膜中，形成用于基因组传递的 DNA 排出体。在这里，我们描述了重组 gp14、gp15 和 gp16 的纯化和表征。该方案用于 T7 周质隧道的高分辨率冷冻电镜结构分析，并且可以适应研究来自其他噬菌体的排出蛋白。有关该方案的使用和执行的完整详细信息，请参阅 Swanson 等人。（2021 年）。
