Detection of the induction of Salmonella enterotoxin gene expression by contact with epithelial cells with RT-PCR

FEMS Microbiol Lett. 1997 Jan 15;146(2):175-9. doi: 10.1111/j.1574-6968.1997.tb10189.x.

Abstract

All strains of Salmonella enterica investigated were found to carry the Salmonella enterotoxin gene (stn) as determined by PCR and hybridization studies. However, when using CHO-K1 cells for testing the toxicity of the strains, not all strains showed a toxic effect (cell elongation) on the cells or did so only at a low level. The cultivation of Salmonella in contact with epithelial cells (IEC-6) led to an increase in the production of toxin. The stn gene expression was detectable with the help of the RT-PCR after 3 h of incubation. The RNA of the strains was isolated, transcribed into cDNA (with MMLV-reverse transcriptase) and amplified using PCR. The PCR products were separated electrophoretically using a polyacrylamide gel and detected by silver staining.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • CHO Cells / microbiology
  • Chlorocebus aethiops
  • Cricetinae
  • DNA Primers
  • DNA, Complementary
  • Enterotoxins / genetics*
  • Epithelium / microbiology
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial / physiology
  • Intestines / cytology
  • Polymerase Chain Reaction
  • Rats
  • Salmonella enteritidis / genetics*
  • Vero Cells / microbiology

Substances

  • Bacterial Proteins
  • DNA Primers
  • DNA, Complementary
  • Enterotoxins
  • enterotoxin, Salmonella