Location of the enterotoxin gene from Salmonella typhimurium and characterization of the gene products.
The enterotoxin gene (stn) in Salmonella typhimurium (Q1 strain) was confined to an 800 bp ClaI-EcoRI genomic DNA fragment (pCE3) that coded for two polypeptides (25 and 12 kDa) under the control of the T7 RNA polymerase/promoter system. The appearance of the 25 kDa protein corresponded to the enterotoxic activity, as determined by elongation of Chinese hamster ovary (CHO) cells, fluid accumulation in rabbit intestinal loops, and altered vascular permeability in rabbit skin. The stn gene products (STN) caused an elevation of intracellular cAMP in CHO cells. These values were at control levels in stn mutants devoid of enterotoxicity, and the 25-kDa protein concurrently disappeared. The biological activity of the heat-labile enterotoxin was blocked by GM1 ganglioside and neutralized by affinity-purified antibodies made against cholera toxin. The 12 kDa protein however was not correlated with an enterotoxic response.
Department of Microbiology, University of Texas Medical Branch, Galveston 77555-1051.
P
P
Chary
R
R
Prasad
A K
AK
Chopra
J W
JW
Peterson
D000818:Animals; D001681:Biological Assay; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004768:Enterotoxins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D008970:Molecular Weight; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011817:Rabbits; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000633:toxicity; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000235:genetics*; D017384:Sequence Deletion; D012867:Skin / Q000187:drug effects / Q000378:metabolism / Q000473:pathology
D004269:DNA, Bacterial; D004768:Enterotoxins; D011994:Recombinant Proteins; C082864:enterotoxin, Salmonella; D000242:Cyclic AMP