Nucleotide sequence and characterization of peb4A encoding an antigenic protein in Campylobacter jejuni.
The 29-kDa protein PEB4, a major antigen of Campylobacter jejuni, is present in all C. jejuni strains tested and elicits an antibody response in infected patients. By screening a lambda gt11 library of chromosomal DNA fragments of C. jejuni strain 81-176 in Escherichia coli Y1090 cells with antibody raised against purified PEB4, a recombinant phage with a 2-kb insert expressing an immunoreactive protein of 29 kDa was isolated. DNA sequence analysis revealed that the insert contains two complete open reading frames ORF-A and ORF-B. ORF-A (peb4A) encodes a 273-residue protein with a calculated molecular mass of 30,460 daltons. The deduced amino acid sequence, composition and pl of the recombinant mature protein are similar to those determined for purified PEB4. The first 21 residues resemble a signal peptide. Gene bank searches indicated 33.7% identity with protein export protein PrsA of Bacillus subtilis and 23.8% identity with protease maturation protein precursor PrtM of Lactococcus lactis. PCR experiments indicate that peb4A is highly conserved among C. jejuni strains. ORF-B begins 2 bp after the last codon of peb4A and encodes a putative protein of 353 residues with 63.4% identity with E. coli fructose 1,6-biphosphate aldolase. The sequence arrangement suggests that these two genes form an operon.
Burucoa
C
C
Laboratoire de Microbiologie A, CHU Poitiers, France.
Frémaux
C
C
Pei
Z
Z
Tummuru
M
M
Blaser
M J
MJ
Cenatiempo
Y
Y
Fauchère
J L
JL
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D016123:Campylobacter jejuni / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D004269:DNA, Bacterial / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005634:Fructose-Bisphosphate Aldolase / Q000737:chemistry / Q000235:genetics; D005784:Gene Amplification; D015151:Immunoblotting; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics*; D015183:Restriction Mapping
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005634:Fructose-Bisphosphate Aldolase