Cloning and expression of the Campylobacter jejuni lon gene detected by RNA arbitrarily primed PCR

FEMS Microbiol Lett. 1998 Aug 15;165(2):329-34. doi: 10.1111/j.1574-6968.1998.tb13165.x.

Abstract

Fingerprinting of RNA by arbitrarily primed PCR was used to identify a heat-inducible gene in Campylobacter jejuni. Comparing RNA fingerprints from C. jejuni cells before and after 20 min of heat shock at 48 degrees C, a differentially amplified PCR product was identified which displayed a high degree of homology to bacterial lon genes. By screening C. jejuni genomic libraries, the entire lon gene was cloned and sequenced. It encodes a protein of 791 amino acids with a calculated molecular mass of 90.2 kDa. Alignment of the Lon amino acid sequence with that of other bacterial species revealed an overall identity of up to 56.6% (Helicobacter pylori). Northern and RNA dot blot experiments confirmed heat induction of the C. jejuni lon gene, revealing a maximum 6-8-fold increase in the level of specific mRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Dependent Proteases
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Campylobacter jejuni / enzymology*
  • Campylobacter jejuni / genetics
  • Cloning, Molecular
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial*
  • Heat-Shock Proteins / chemistry
  • Heat-Shock Proteins / genetics*
  • Heat-Shock Proteins / metabolism*
  • Heat-Shock Response
  • Humans
  • Molecular Sequence Data
  • Plasmids
  • Polymerase Chain Reaction / methods*
  • RNA, Bacterial / analysis
  • RNA, Bacterial / isolation & purification
  • Restriction Mapping
  • Sequence Analysis, DNA
  • Serine Endopeptidases / chemistry
  • Serine Endopeptidases / genetics*
  • Serine Endopeptidases / metabolism*
  • Transcription, Genetic

Substances

  • Heat-Shock Proteins
  • RNA, Bacterial
  • ATP-Dependent Proteases
  • Serine Endopeptidases

Associated data

  • GENBANK/Y17166