Identification of multicopy suppressors of the pcnB plasmid copy number defect in Escherichia coli

N. Sarkar, G. j. Cao, C. Jain

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Plasmids containing a ColE1 origin of replication are widely used for cloning purposes in Escherichia coli. Among the host factors that affect the copy number of ColE1 plasmids is the E. coli protein poly(A) polymerase I (PAP I), which regulates the intracellular level of RNA I, a ColE1-encoded negative regulator of plasmid replication. In strains that lack PAP I, RNA I levels are elevated, resulting in reduced levels of ColE1 plasmids in the cell. PAP I is encoded by the gene pcnB. We devised a genetic approach, based on the identification of multicopy suppressor clones, to identify transacting factors that can help offset the ColE1 plasmid copy number defect in a pcnB- genetic background. Using this strategy, we identified suppressors that mapped to two regions of the E. coli chromosome. The suppressor activity of one of the chromosomal regions was localized to the rssB gene, a response regulator gene known to be involved in the turnover of the stationary-phase sigma factor, RpoS. The second suppressor maps to min 55.4 of the E. coli chromosome, and the factor responsible for the suppressor activity appears to be a novel RNA or protein.

Original languageEnglish (US)
Pages (from-to)62-69
Number of pages8
JournalMolecular Genetics and Genomics
Volume268
Issue number1
DOIs
StatePublished - Oct 10 2002

Keywords

  • Antisense RNA
  • ColE1 plasmid
  • Plasmid copy number
  • Poly(A) polymerase

ASJC Scopus subject areas

  • Genetics

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