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Methods Mol Biol. 2014;1144:265-81. doi: 10.1007/978-1-4939-0428-0_18.

Cryo-EM techniques to resolve the structure of HSV-1 capsid-associated components.

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1
Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA.

Abstract

Electron cryo-microscopy has become a routine technique to determine the structure of biochemically purified herpes simplex virus capsid particles. This chapter describes the procedures of specimen preparation by cryopreservation; low dose and low temperature imaging in an electron cryo-microscope; and data processing for reconstruction. This methodology has yielded subnanometer resolution structures of the icosahedral capsid shell where α-helices and β-sheets of individual subunits can be recognized. A relaxation of the symmetry in the reconstruction steps allows us to resolve the DNA packaging protein located at one of the 12 vertices in the capsid.

PMID:
24671690
PMCID:
PMC4370171
DOI:
10.1007/978-1-4939-0428-0_18
[Indexed for MEDLINE]
Free PMC Article
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