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

• Published in: FEMS microbiology letters 1997-01, Vol.146 (2), p.175-179
• Main Authors: Dinjus, Ute, Hänel, Ingrid, Müller, W, Bauerfeind, R, Helmuth, R
• Format: Article
• Language: English
• Subjects: Animals; Bacterial Proteins - genetics; Cercopithecus aethiops; CHO Cells - microbiology; Complementary DNA; Cricetinae; Cultivation; DNA Primers; DNA, Complementary; Elongation; Enterotoxin; Enterotoxins - genetics; Epithelial cell; Epithelial cells; Epithelium - microbiology; Escherichia coli - genetics; Gene expression; Gene Expression Regulation, Bacterial - physiology; Hybridization; Intestines - cytology; Low level; Microbiology; Polyacrylamide; Polymerase Chain Reaction; Rats; Ribonucleic acid; RNA; RNA-directed DNA polymerase; Salmonella; Salmonella enterica; Salmonella enteritidis - genetics; Toxicity; Toxicity testing; Toxins; Vero Cells - microbiology
• ISSN: 0378-1097; 1574-6968
• 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.