A GapC chimera retains the properties of the Streptococcus uberis wild-type GapC protein.
The GapC products of Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis share considerable homology at the DNA and amino acid levels. The high similarity at the protein level suggests that one GapC protein might be used as a single antigen to protect dairy cows against infections with the contagious S. agalactiae and the environmental S. dysgalactiae and S. uberis strains. Despite their similarities, immunization with the S. dysgalactiae GapC did not protect dairy cows from a challenge with S. uberis, suggesting the presence of regions in GapC that are involved in species-specific protection. To produce a single antigen that can be used to protect against all streptococcal mastitis infections, we constructed a GapC chimeric protein using the S. uberis GapC product as the backbone followed by non-conserved peptide regions from the S. agalactiae and S. dysgalactiae GapC proteins. We report that the chimeric GapC protein retains the enzymatic activity of the S. uberis GapC protein. In addition, we fused the chimera to the OmpF and LipoF transport sequences of Escherichia coli and the GapC chimeras were present in membrane fractions of E. coli. These extracts could be the basis of an antigen preparation for use in mastitis vaccines.
Perez-Casal
Jose
J
Vaccine and Infectious Disease Organization, University of Saskatchewan, 120 Veterinary Road, Saskatoon, Saskatchewan, Canada S7N 5E3. perezcasal@sask.usask.ca
Prysliak
Tracy
T
Potter
Andrew A
AA
D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D002417:Cattle; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D007700:Kinetics; D008414:Mastitis, Bovine / Q000517:prevention & control; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry; D010446:Peptide Fragments / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D022541:Streptococcal Vaccines / Q000235:genetics* / Q000276:immunology; D013291:Streptococcus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003470:Culture Media; C469817:GapC protein, Streptococcus; D009841:Oligonucleotides; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D022541:Streptococcal Vaccines; C068142:glyceraldehyde 3-phosphate dehydrogenase (304-313); D006639:Histidine; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases