Ribosome Profiling of Vaccinia Virus-Infected Cells

Methods Mol Biol. 2019:2023:171-188. doi: 10.1007/978-1-4939-9593-6_11.

Abstract

Ribosome profiling is a method that determines genome-wide mRNA translation through measuring ribosome-protected mRNA fragments by deep sequencing. This method can be used to quantify gene expression at the translational level and precisely pinpoint ribosome loading onto mRNA with codon-level resolution. Genome-wide regulation of mRNA translation can also be determined if RNA-Sequencing (RNA-Seq) is carried out in parallel. Here, we describe a protocol for simultaneously performing ribosome profiling and RNA-Seq in cells infected with vaccinia virus.

Keywords: Gene expression; Poxvirus; RNA-Seq; Ribosome profiling; Translation; Vaccinia virus.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Protein Biosynthesis / genetics
  • RNA, Messenger / genetics
  • Ribosomes / genetics
  • Ribosomes / metabolism*
  • Vaccinia virus / genetics*

Substances

  • RNA, Messenger