Cloning and sequencing of the gene encoding Vibrio cholerae O1 fimbrial subunit (fimbrillin).
The gene encoding an 18 kDa fimbrial subunit of Vibrio cholerae O1 was identified in a fimbriate strain Bgd17. Mixed oligoprimers were prepared based on the amino acid sequence of the N-terminus and that from a cyanogen bromide-cleaved fragment of the fimbrillin. A PCR-amplified 185 bp DNA fragment was sequenced. This 185 bp fragment was further extended to 540 bp to 3' and 5' termini by RNA-PCR using a primer containing a random hexamer at its 3' end. This fragment did not contain the stop codons. It was further extended by a gene walking method using Eco RI cassette and its primers. Finally a 660 bp fragment was obtained and sequenced. This fragment contained the complete open reading frame of the structural subunit of the fimbriae, composed of 169 amino acids with a molecular mass of 17435.65 and a leader sequence of 6 or 9 amino acids. The deduced amino acid sequence of the polypeptide encoded by the gene, designated fimA, displayed a highly conserved sequence of MKXXXGFTLI EL of type 4 fimbriae.
Ehara
M
M
Department of Bacteriology, Nagasaki University, Japan.
Ichinose
Y
Y
Hirayama
T
T
Kurazono
H
H
Hidaka
Y
Y
Morita
K
K
Igarashi
A
A
Shimodori
S
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D015342:DNA Probes; D036781:Fimbriae Proteins; D008969:Molecular Sequence Data; D014734:Vibrio cholerae / Q000235:genetics* / Q000648:ultrastructure
D001426:Bacterial Proteins; D015342:DNA Probes; C063055:fimbrillin; D036781:Fimbriae Proteins