[Cloning and expression of Vibrio cholerae zonula occludens toxin (zot) gene in Escherichia coli].
Two recombinant plasmids containing the cloned PCR-amplifled Vibrio cholerae zonula occludens toxin (zot) gene was constructed in orientation providing its transcription from lac-promoter. One of them contained also its own zot promoter. The third plasmid was obtained by subcloning a Vibrio cholerae DNA fragment including intact zot and ace (accessory cholera enterotoxin) genes. The expression levels of the cloned genes in Escherichia coli varied depending on a promoter type, host strain and culture conditions. The human intestinal cell line CaCo2 appeared to be a suitable model for assessing the biological activity of toxin preparations. The product of zot gene possessed a marked activity in respect to CaCo2 in spite of the lack of the molecule cleavage and transport of its toxic C-terminal part from alien host cells into the culture media. The constructed recombinant plasmids can be used as a source of molecular hybridization probes; and E.col transformants carrying those plasmids can serve in zot purification both for the scientific and practical purposes.
Pisanov
R V
RV
Goncharov
E K
EK
Monakhova
E V
EV
Alekseeva
L P
LP
Mishan'kin
B N
BN
D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000235:genetics; D018938:Caco-2 Cells; D002772:Cholera Toxin / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004731:Endotoxins; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D006801:Humans; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D014734:Vibrio cholerae / Q000235:genetics*
C081530:Ace toxin; D004731:Endotoxins; D011994:Recombinant Proteins; C077728:zonula occludens toxin, Vibrio cholerae; D002772:Cholera Toxin; D000251:Adenosine Triphosphatases