Recombinant expression of a truncated capsid protein of beak and feather disease virus and its application in serological tests.
Beak and feather disease virus (BFDV) causes severe disease characterized by irreversible feather disorders and severe immunosuppression in many psittacine species. BFDV cannot be propagated in tissue or cell cultures, rendering virus propagation and thus diagnosis rather difficult. To develop reliable diagnostic methods, the region encoding the BFDV capsid protein C1 was cloned from an infected sulphur-crested cockatoo (Cacatua galerita). Phylogenetic analysis showed this gene had 76.3 to 83.2% amino acid identity to published sequences. No protein was detected after induction of full-length C1 expression in Escherichia coli. However, deletion of an amino-terminal arginine-rich sequence facilitated expression. C1(39-244)-His, a polyhistidine-tailed variant of this protein, was purified and used for immunization of chickens. The immune sera detected C1 with an apparent molecular weight of 27 kDa in western blots of organ homogenates of BFDV-infected birds. Using C1(39-244)-His as antigen, 11 psittacine sera were tested for the presence of BFDV-specific antibodies by enzyme-linked immunosorbent assay and immunoblotting. The results obtained correlated well with the BFDV-specific haemagglutination inhibition activity of the sera, suggesting C1(39-244)-His has value as a recombinant antigen for BFDV-specific serological tests.
Johne
Reimar
R
Institute for Virology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 29, D-04103 Leipzig, Germany. johne@vetmed.uni-leipzig.de
Raue
Rüdiger
R
Grund
Christian
C
Kaleta
Erhard F
EF
Müller
Hermann
H
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001483:Base Sequence; D001715:Bird Diseases / Q000175:diagnosis / Q000276:immunology / Q000821:virology*; D001717:Birds; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D018173:Circoviridae Infections / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary*; D017925:Circovirus / Q000235:genetics* / Q000276:immunology; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D040461:Gene Components; D007106:Immune Sera / Q000276:immunology; D007114:Immunization / Q000662:veterinary*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D000914:Antibodies, Viral; D036022:Capsid Proteins; D017931:DNA Primers; D007106:Immune Sera; D011994:Recombinant Proteins