Open AccessDetection of the induction of Salmonella enterotoxin gene expression by contact with epithelial cells with RT‐PCR
Author(s) -
Dinjus Ute,
Hänel Ingrid,
Müller W,
Bauerfeind R,
Helmuth R
Publication year - 1997
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1997.tb10189.x
Subject(s) - salmonella , enterotoxin , biology , microbiology and biotechnology , complementary dna , salmonella enterica , gene expression , gene , reverse transcriptase , polymerase chain reaction , incubation , toxin , real time polymerase chain reaction , bacteria , escherichia coli , biochemistry , genetics
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.