Systematic chromosomal deletion of bacterial ribosomal protein genes

Shinichiro Shoji, Corey M. Dambacher, Zahra Shajani, James R. Williamson, Peter G. Schultz

Research output: Contribution to journalArticlepeer-review

69 Scopus citations

Abstract

Detailed studies of ribosomal proteins (RPs), essential components of the protein biosynthetic machinery, have been hampered by the lack of readily accessible chromosomal deletions of the corresponding genes. Here, we report the systematic genomic deletion of 41 individual RP genes in Escherichia coli, which are not included in the Keio collection. Chromosomal copies of these genes were replaced by an antibiotic resistance gene in the presence of an inducible, easy-to-exchange plasmid-born allele. Using this knockout collection, we found nine RPs (L15, L21, L24, L27, L29, L30, L34, S9, and S17) nonessential for survival under induction conditions at various temperatures. Taken together with previous results, this analysis revealed that 22 of the 54 E. coli RP genes can be individually deleted from the genome. These strains also allow expression of truncated protein variants to probe the importance of RNA-protein interactions in functional sites of the ribosome. This set of strains should enhance in vivo studies of ribosome assembly/function and may ultimately allow systematic substitution of RPs with RNA.

Original languageEnglish (US)
Pages (from-to)751-761
Number of pages11
JournalJournal of molecular biology
Volume413
Issue number4
DOIs
StatePublished - Nov 4 2011
Externally publishedYes

Keywords

  • protein engineering
  • recombineering
  • ribonucleoprotein complex
  • ribosome
  • translation

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Fingerprint

Dive into the research topics of 'Systematic chromosomal deletion of bacterial ribosomal protein genes'. Together they form a unique fingerprint.

Cite this