Detection of a novel Borna disease virus-encoded 10 kDa protein in infected cells and tissues.
Borna disease (BD) is a transmissible, progressive polioencephalomyelitis primarily of horses and sheep. The genomes of two cell-adapted strains of Borna disease virus (BDV), the aetiological agent of BD, have been cloned and sequenced. According to the structural characterization achieved so far, BDV contains a non-segmented negative-sense 8.9 kb single-stranded RNA genome. In this paper we report the expression, purification and intracellular tracing of a novel non-glycosylated BDV-specific protein with a molecular mass of approximately 10 kDa (BDV p10 protein). The successful isolation of the corresponding mRNA from infected cells, amplification of the genetic region by RT-PCR and its efficient expression as a glutathione S-transferase (GST) fusion protein demonstrated that antibodies specific for the BDV p10 protein are induced in infected animals. In addition, we have produced monospecific antisera against the GST-p10 fusion protein in rabbits. This monospecific antiserum recognized the BDV p10 protein in brain cells of naturally and experimentally infected animals as well as in persistently BDV-infected cells. Antibody-mediated affinity-chromatography using the anti-p10 serum could successfully be applied to purify a ca. 10 kDa antigen from infected animal cells to such an extent that glycosylation of this component could be ruled out.
Wehner
T
T
Institut für Virologie, Giessen, Germany.
Ruppert
A
A
Herden
C
C
Frese
K
K
Becht
H
H
Richt
J A
JA
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D001483:Base Sequence; D001890:Borna Disease / Q000821:virology*; D001891:Borna disease virus / Q000737:chemistry*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004285:Dogs; D005814:Genes, Viral; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D015678:Viral Structural Proteins / Q000235:genetics
D000914:Antibodies, Viral; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins