Detection of the induction of Salmonella enterotoxin gene expression by contact with epithelial cells with RT-PCR.
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.
Dinjus
U
U
Federal Institute for Health Protection of Consumers and Veterinary Medicine, Division 4, Bacterial Epizootics and Zoonoses Control, Jena, Germany.
Hänel
I
I
Müller
W
W
Bauerfeind
R
R
Helmuth
R
R
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D016466:CHO Cells / Q000382:microbiology; D002522:Chlorocebus aethiops; D006224:Cricetinae; D017931:DNA Primers; D018076:DNA, Complementary; D004768:Enterotoxins / Q000235:genetics*; D004848:Epithelium / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D007422:Intestines / Q000166:cytology; D016133:Polymerase Chain Reaction; D051381:Rats; D012477:Salmonella enteritidis / Q000235:genetics*; D014709:Vero Cells / Q000382:microbiology
D001426:Bacterial Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D004768:Enterotoxins; C082864:enterotoxin, Salmonella