Marek's disease virus protein kinase gene identified within the short unique region of the viral genome is not essential for viral replication in cell culture and vaccine-induced immunity in chickens.
The open reading frame (ORF) of 1206 bp within the short unique region (Us) of Marek's disease virus type 1 (MDV1) shows significant homology with the herpes simplex virus type 1 US3 gene encoding protein kinase (PK). The lacZ gene of Escherichia coli was inserted within the ORF, designated MDV1-US3, of MDV1 K544 strain DNA by homologous recombination. The plaque-purified recombinant MDV1 stably expressed the beta-galactosidase encoded by the inserted lacZ gene in infected cells and replicated well as the parental K544 strain. Antibodies against both MDV1 antigen and beta-galactosidase were detected in the sera of chickens immunized with recombinant MDV1. Chickens vaccinated with the recombinant MDV1 were protected from challenge with virulent MDV1. The MDV1 US3 gene expressed by a baculovirus vector encoded a 44-kDa protein. Mouse antisera against the 44-kDa protein reacted with two proteins of 44 and 45 kDa in extracts of cells infected with MDV1 but not with MDV types 2 or 3. The PK activity was detected in immune complexes of the anti-44-kDa sera with extracts of cells infected with MDV1 but not with the recombinant MDV1. Thus, PK encoded from the MDV1-US3 is not essential for virus replication in cell culture and vaccine-induced immunity.
Chemo-Sero-Therapeutic Research Institute, Kikuchi Research Center, Kumamoto, Japan.
M
M
Sakaguchi
T
T
Urakawa
Y
Y
Hirayama
N
N
Miki
M
M
Yamamoto
G S
GS
Zhu
K
K
Hirai
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D004279:DNA, Viral; D004285:Dogs; D005822:Genetic Vectors; D016679:Genome, Viral; D008381:Herpesvirus 2, Gallid / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D007763:Lac Operon; D008380:Marek Disease / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D014765:Viral Vaccines / Q000276:immunology*; D014779:Virus Replication / Q000235:genetics
D004274:DNA, Recombinant; D004279:DNA, Viral; D014765:Viral Vaccines; D011494:Protein Kinases