Characterization of detergent-soluble proteins of Corynebacterium pseudotuberculosis.
Whole cells and culture supernatant of isolates of Corynebacterium pseudotuberculosis were studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting (immunoblotting). SDS-PAGE analysis of detergent-solubilized whole cells revealed more than 20 bands in silver-stained gels. However, the SDS-soluble proteins that are present in all the isolates of the bacterium can be separated into four groups, as follows, (i) high molecular mass proteins that are greater than 119 kDa, (ii) a set of three proteins with molecular mass of 84, 64 and 58 kDa, (iii) a doublet consisting of proteins of molecular mass 33 to 30 kDa, and (iv) low molecular mass proteins of 25 to 20 kDa. SDS-PAGE analysis of ammonium sulphate concentrated culture supernatant demonstrated more than seven bands in silver-stained gels ranging in molecular mass from 64-14 kDa. Sera from goats with C. pseudotuberculosis-induced disease were used to probe immunoblots of electrophoresed SDS-soluble proteins. Ten or more SDS-soluble proteins from whole cells, ranging in molecular mass from 119-20 kDa were recognized by antibodies in sera of naturally infected goats. These sera also recognized up to five molecules ranging from 64-30 kDa, on immunoblots of ammonium sulfate concentrated culture supernatant.
Braithwaite
C E
CE
Cooperative Agricultural Research Center (CARC), Prairie View A&M University, TX 77446-2886.
Smith
E E
EE
Songer
J G
JG
Reine
A H
AH
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western / Q000662:veterinary; D003354:Corynebacterium Infections / Q000276:immunology; D016925:Corynebacterium pseudotuberculosis / Q000737:chemistry* / Q000276:immunology; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D015511:Goat Diseases / Q000276:immunology; D006041:Goats; D006736:Horses; D007106:Immune Sera / Q000276:immunology; D008970:Molecular Weight; D012756:Sheep; D016622:Silver Staining / Q000662:veterinary; D013047:Specific Pathogen-Free Organisms
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003902:Detergents; D007106:Immune Sera