Cloning of the fliI gene from Rhodobacter sphaeroides WS8 by analysis of a transposon mutant with impaired motility

FEMS Microbiol Lett. 1996 Aug 15;142(1):111-6. doi: 10.1111/j.1574-6968.1996.tb08416.x.

Abstract

A transposon mutant of Rhodobacter sphaeroides WS8 was isolated that showed reduced swarming on soft agar plates. Liquid cultures of this mutant (M18) showed a low percentage of motile swimming cells in mid-exponential phase and a low level of extracellular flagellin protein by Western blotting. M18 was complemented by a clone from a library of R. sphaeroides WS8 DNA, and restriction mapping of the site of TnphoA insertion in the mutant, coupled with DNA sequencing, showed that it had a defect in the fliI gene. To determine if a partly functional fliI gene was giving the low-motility phenotype of M18, a drug resistance omega cartridge was inserted into the gene to give a complete null mutant. This null strain also produced a low percentage of motile cells. Possible reasons for this apparent fliI-independent flagellar formation are discussed.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins*
  • DNA Transposable Elements
  • Flagella / physiology
  • Genes, Bacterial*
  • Molecular Sequence Data
  • Mutation
  • Proteins / genetics*
  • Proton-Translocating ATPases*
  • Restriction Mapping
  • Rhodobacter sphaeroides / genetics*
  • Rhodobacter sphaeroides / physiology
  • Salmonella typhimurium / genetics
  • Sequence Homology, Amino Acid
  • Xanthomonas campestris / genetics

Substances

  • Bacterial Proteins
  • DNA Transposable Elements
  • Proteins
  • fliI protein, bacteria
  • Proton-Translocating ATPases

Associated data

  • GENBANK/X97201