Dinjus U, Hänel I, Müller W, Bauerfeind R, Helmuth R
Federal Institute for Health Protection of Consumers and Veterinary Medicine, Division 4, Bacterial Epizootics and Zoonoses Control, Jena, Germany.
FEMS Microbiol Lett. 1997 Jan 15;146(2):175-9. doi: 10.1111/j.1574-6968.1997.tb10189.x.
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.
通过聚合酶链反应（PCR）和杂交研究确定，所有被研究的肠炎沙门氏菌菌株均携带沙门氏菌肠毒素基因（stn）。然而，当使用中国仓鼠卵巢细胞系-K1（CHO-K1）细胞检测这些菌株的毒性时，并非所有菌株都对细胞产生毒性作用（细胞伸长），或者仅在低水平上产生这种作用。将沙门氏菌与上皮细胞（IEC-6）接触培养会导致毒素产生增加。孵育3小时后，借助逆转录聚合酶链反应（RT-PCR）可检测到stn基因表达。分离菌株的RNA，使用莫洛尼氏鼠白血病病毒逆转录酶（MMLV-逆转录酶）将其转录为互补DNA（cDNA），并通过PCR进行扩增。使用聚丙烯酰胺凝胶对PCR产物进行电泳分离，并通过银染进行检测。
