Expression and purification of phage T7 ejection proteins for cryo-EM analysis

Authors

Nicholas A. Swanson, Thomas Jefferson UniversityFollow
Ravi K Lokareddy, Thomas Jefferson UniversityFollow
Fenglin Li, Thomas Jefferson UniversityFollow
Chun-Feng Hou, Thomas Jefferson UniversityFollow
Mikhail Pavlenok, University of Alabama at Birmingham
Michael Niederweis, University of Alabama at Birmingham
Gino Cingolani, Thomas Jefferson UniversityFollow

Document Type

Article

Publication Date

11-16-2021

Comments

This article is the author’s final published version in STAR Protocols, Volume 2, Issue 4, November 2021, Article number 100960

The published version is available at https://doi.org/10.1016/j.xpro.2021.100960. Copyright © Swanson et al.

Abstract

Bacteriophages of the Podoviridae 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.

Recommended Citation

Swanson, Nicholas A.; Lokareddy, Ravi K; Li, Fenglin; Hou, Chun-Feng; Pavlenok, Mikhail; Niederweis, Michael; and Cingolani, Gino, "Expression and purification of phage T7 ejection proteins for cryo-EM analysis" (2021). Department of Biochemistry and Molecular Biology Faculty Papers. Paper 199.
https://jdc.jefferson.edu/bmpfp/199

Creative Commons License

This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

PubMed ID

34825220

Language

English