Molecular cloning and expression of Salmonella typhi flagellin: characterization of 52 kDa specific antigen of S. typhi.
We previously reported monoclonal antibodies (MAbs) specific to S. typhi 52 kDa antigen which do not cross react with related protein antigens from 11 bacteria causing enteric fever and enteric fever-like illness. Using the combination of these specific MAbs and recombinant DNA technology, expression plasmids containing the antigen gene producing substantial amount of the S. typhi protein antigen have been established. Plasmid pSKM-T7 containing the specific 52 kDa antigen gene was cloned and the antigen expressed was detectable by immunoblotting using specific mAbs. The complete nucleotide sequence of this gene was compared with other bacterial sequences and found to be highly homologous with the flagellin gene H1-d of S. muenchen except in the hypervariable region in the central portion. The specific 52 kDa antigen of S. typhi detected by our MAbs is thus a flagellin.
Sukosol
T
T
Department of Immunology, Faculty of Medicine Siriraj Hospital, Bangkok, Thailand.
Sarasombath
S
S
Mongkolsuk
S
S
Songsivilai
S
S
Chaiyaroj
S
S
Pongsunk
S
S
Ekpo
P
P
D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005408:Flagellin / Q000235:genetics* / Q000276:immunology; D015964:Gene Expression Regulation, Bacterial; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010957:Plasmids; D012485:Salmonella typhi / Q000235:genetics* / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D004269:DNA, Bacterial; D005408:Flagellin