Genome-Wide Quantitation of Protein Synthesis Rates in Bacteria
Name
nihms-1040364.pdf
Description
Accepted version
Size
1.38 MB
Format
Adobe PDF
Checksum (MD5)
d1e6d91e2cb6df965b870a65e6330b70
Author(s)
Johnson, Grace E.
Li, Gene-Wei
Date Issued
December 2018
Journal
Methods in enzymology
Publisher
Elsevier
Citation
Johnson, Grace E. and Gene-Wei Li. “Genome-Wide Quantitation of Protein Synthesis Rates in Bacteria.” Methods in enzymology 612 (2018): 225-249 © 2018 The Author(s)
Version
Author's final manuscript
Abstract
Bacteria produce different amounts of their proteins in response to different conditions. The ability to accurately quantitate the rates of protein synthesis across the genome is an important step toward understanding both underlying regulation and bacterial physiology at a systems level. Ribosome profiling (deep sequencing of ribosome-protected mRNA fragments) enables accurate and high-throughput measurement of such synthesis rates. Ribosomes protect RNAs from nuclease digestion; thus, by collecting and sequencing protected footprints, one can obtain information on the position of every ribosome at the time of cell collection. Assuming ribosomes go on to translate full-length proteins, the density of ribosomes across an ORF can be used to determine protein synthesis rates. Here we outline a step-by-step protocol and discuss the steps where variability and bias may be introduced, including ways to minimize it.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/bs.mie.2018.08.031
