title: [Cloning of the zot gene of Vibrio cholerae and its expression in Escherichia coli].
abstract: The zot gene encoding Zonula occludens toxin was amplified from classic Vibrio cholerae genomic DNA by PCR. The result of sequencing indicated that zot gene encodes 399 amino acid residues. The sequence of zot gene was a little bit different from that of reported including 14 nucleotides and four amino acid residues. The expression plasmid pET-ZOT was constructed by inserting zot gene into plasmid pET-28a(+) containing the T7 promoter. The expression plasmid was induced into E. coli BL21 (DE3) and expression strain BLZOT was selected. SDS-PAGE analysis revealed that the ZOT protein was expressed and accumulated up to above 15% of bacterial soluble protein after induced by IPTG. A protein of 47 kD was expressed as including body. Western blot analysis revealed that the expressed protein was ZOT.
authors.affiliation: Shanghai Institute of Biochemistry, Chinese Academy of Sciences, Shanghai 200031.
authors.forename: Z Y
authors.initials: ZY
authors.lastname: He
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Chen
authors.forename: D N
authors.initials: DN
authors.lastname: Wang
authors.forename: G Z
authors.initials: GZ
authors.lastname: Yang
authors.forename: W J
authors.initials: WJ
authors.lastname: Zhang
authors.forename: X F
authors.initials: XF
authors.lastname: Wu
mesh_terms: D001483:Base Sequence; D002772:Cholera Toxin / Q000235:genetics*; D003001:Cloning, Molecular; D004731:Endotoxins; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014734:Vibrio cholerae / Q000235:genetics*
chemical_list: D004731:Endotoxins; D011994:Recombinant Proteins; C077728:zonula occludens toxin, Vibrio cholerae; D002772:Cholera Toxin