Monitoring Bacterial Translation Rates Genome-Wide

Methods Mol Biol. 2021:2252:3-26. doi: 10.1007/978-1-0716-1150-0_1.

Abstract

Modern DNA sequencing technologies have allowed for the sequencing of tens of thousands of bacterial genomes. While this explosion of information has brought about new insights into the diversity of the prokaryotic world, much less is known of the identity of proteins encoded within these genomes, as well as their rates of production. The advent of ribosome profiling, or the deep sequencing of ribosome-protected footprints, has recently enabled the systematic evaluation of every protein-coding region in a given experimental condition, the rates of protein production for each gene, and the variability in translation rates across each message. Here, I provide an update to the bacterial ribosome profiling approach, with a particular emphasis on a simplified strategy to reduce cloning time.

Keywords: Bacterial ribosome profiling; Bacterial translation; Ribo-seq; Ribosome profiling.

MeSH terms

  • Bacteria / genetics
  • Bacteria / metabolism*
  • Bacterial Proteins / metabolism
  • High-Throughput Nucleotide Sequencing
  • Protein Biosynthesis
  • RNA, Bacterial / genetics
  • RNA, Messenger / genetics*
  • Ribosomes / metabolism*
  • Sequence Analysis, RNA

Substances

  • Bacterial Proteins
  • RNA, Bacterial
  • RNA, Messenger