Identification of a fljA gene on a linear plasmid as the repressor gene of fliC in Salmonella enterica serovar Typhi

Microbiol Immunol. 2009 Apr;53(4):191-7. doi: 10.1111/j.1348-0421.2009.00106.x.

Abstract

Salmonella enterica serovar Typhi z66-positive strain contains an fljBA-like operon on a linear plasmid and the fliC gene on the chromosome encoding d or j antigen. The fljB-like gene has been identified as the gene encoding z66 antigen. To investigate the function of the fljA-like gene in S. enterica serovar Typhi, a z66-positive wild-type strain GIFU10007 and its phase variation j positive strain 007j+ were used in the present study. After deletion of the fljA-like gene, the wild-type strain that only expressed FljB:z66 could express both FliC:j and FljB:z66 at the same time. A recombinant plasmid, pBADfljA, containing the fljA-like gene was prepared. SDS-PAGE and western blot analysis of secreted proteins showed that when the strain 007j+ was transformed by the recombinant plasmid, pBADfljA, the expression of FliC:j was inhibited. The fliC::lacZ fusion of the strain 007j+ was constructed. Beta-Galactosidase activity was decreased approximately 100-fold after harboring the plasmid pBADfljA in the fliC::lacZ mutant strain. However, the fliC mRNA level investigated by RT-PCR was decreased only approximately seven-fold in the strain 007j+ after harboring the plasmid pBADfljA. These results demonstrated that thefljA-like gene on a linear plasmid of S. enterica serovar Typhi is a fljA gene, the repressor of fliC gene, and the inhibition by FljA is likely at the post-transcriptional level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / biosynthesis*
  • Artificial Gene Fusion
  • Bacterial Proteins / genetics
  • Bacterial Proteins / physiology*
  • Gene Deletion
  • Gene Expression Profiling
  • Genes, Bacterial*
  • Genes, Reporter
  • Plasmids*
  • Repressor Proteins / genetics
  • Repressor Proteins / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Salmonella typhi / genetics*
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • Antigens, Bacterial
  • Bacterial Proteins
  • FljA protein, Salmonella
  • Repressor Proteins
  • beta-Galactosidase