Expression and immunogenicity of six putative variable surface proteins in Mycoplasma mycoides subsp. mycoides SC.
Variable surface protein Vmm and five Vmm-type proteins from Mycoplasma mycoides subsp. mycoides SC were analysed to determine whether these proteins are expressed in vivo in animals affected by contagious bovine pleuropneumonia (CBPP) and in vitro. Recombinant versions of these proteins were constructed and expressed in Escherichia coli after mutation of the TGA Trp codons to TGG. These proteins were then analysed by dot and Western blotting with sera from CBPP-affected cattle. Furthermore, affinity-purified polyclonal antibodies to the recombinant proteins were used in Western and colony blotting to look for expression of the putative Vmm-type proteins in cultured M. mycoides SC. This study demonstrates that immunoglobulins in CBPP sera recognize all putative Vmm-type proteins tested, indicating that these proteins or their homologues are expressed by mycoplasmas during natural infections. Vmm and one of the putative Vmm-type proteins showed variable expression in vitro.
Hamsten
Carl
C
Department of Proteomics, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden.
Westberg
Joakim
J
Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University, SE-752 36 Uppsala, Sweden.
Bölske
Göran
G
Department of Bacteriology, National Veterinary Institute (SVA), SE-751 89 Uppsala, Sweden.
Ayling
Roger
R
The Mycoplasma Group, Veterinary Laboratories Agency (VLA), Addlestone, Surrey KT15 3NB, UK.
Uhlén
Mathias
M
Department of Proteomics, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden.
Persson
Anja
A
Department of Proteomics, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden.
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology*; D003001:Cloning, Molecular; D007114:Immunization; D015151:Immunoblotting; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D016296:Mutagenesis; D009176:Mycoplasma mycoides / Q000235:genetics / Q000276:immunology*; D011002:Pleuropneumonia, Contagious / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins