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Infect. Immun., Apr 1996, 1233-1239, Vol 64, No. 4
GI Viboud, G Jonson, E Dean-Nystrom and AM Svennerholm
Putative colonization factor PCFO20 was recently identified in an
enterotoxigenic Escherichia coli (ETEC) strain of serogroup O20 isolated
from a child with diarrhea in Argentina. The gene encoding the structural
subunit of PCFO20 fimbriae, fotA, was cloned from strain ARG- 2 in the
expression phage vector lambda ZAP Express. One positive clone, pGV29, that
carried a 3.3-kb fragment was identified on the basis of fimbrillin
production by using a monospecific rabbit anti- PCFO20 serum. Nucleotide
sequencing of a 1.3-kb Sau3A-ClaI fragment of the subclone pGV292
containing the region coding for PCFO20 fimbrillin revealed two open
reading frames of which one was complete. A western blot (immunoblot)
showed that the cloned protein, FotA, migrated like the PCFO20 fimbrial
subunit protein did. Fimbriae were not detected on the surface of E. coli
host bacteria containing pGV292 or pGV29, suggesting that the genes needed
for assembly of PCFO20 fimbriae are lacking in both clones. The fotA gene
encodes a 20,574-Da prefimbrillin protein which contains a 21-amino-acid
signal sequence; the mature protein has a size of 18.1 kDa. The subunit
protein FotA was found to be more homologous to the subunit of porcine 987P
than to any fimbrial subunit produced by human ETEC. Alignments of the
amino acid sequences of the two proteins indicate that they are partly
identical, with an overall similarity of 82%. FotA fimbrillin was shown to
be transported and assembled by the fimbria assembly machinery in porcine
ETEC strain 987. PCFO20 and 987P may have evolved from a common ancestral
gene. They are immunologically related but have affinity for different host
cell receptors, since PCFO20-producing bacteria do not bind to neonatal
piglet enterocytes.
Copyright © 1996, American Society for Microbiology
The structural gene encoding human enterotoxigenic Escherichia coli PCFO20 is homologous to that for porcine 987P
Department of Medical Microbiology and Immunology, Goteborg University, Sweden.
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