Identification of viral proteins encoded by two DNA fragments of herpesvirus of turkeys (HVT).
Herpesvirus of turkeys (HVT) vaccine is used worldwide to immunize chickens against Marek's disease (MD). Polyclonal antiserum directed against one virus cross-reacts with proteins of the other, while only 5% homology at the DNA level was demonstrated between the two viruses. A partial library of HVT DNA fragments ranging from 1.5 to 13.5 kbp in size was constructed in pBR 322. Under stringent conditions of hybridization (low salt concentration, 10% dextran sulfate at 68 degrees C), two of the cloned HVT DNA fragments (13.5 and 11.0 kbp) hybridized to three MDV DNA fragments: BamHI-C, D, and G. The 13.5 kbp fragment detected two transcripts of 1.8 and 3.0 kb in RNA extracted from HVT-infected chicken embryo fibroblasts, while the 11.0 kbp fragment detected three transcripts of 1.6, 2.0, and 3.0 kb in the extracted RNA. Using hybrid selection, specific RNA was isolated by hybridization with the two cloned HVT DNA fragments and then was translated in vitro using rabbit reticulocyte lysate, and the resulting proteins were analyzed by NaDodSO4-PAGE. The RNA selected by the two HVT DNA fragments coded for a 30 kD protein and for several smaller proteins 10-20 kD in size, while the RNA selected by the 11.0 kb HVT DNA fragment was translated into a 40 kD protein. Immunoprecipitation of these in vitro synthesized proteins with hyperimmune anti-HVT chicken serum showed that they were of viral origin.
Levy
H
H
Department of Molecular Virology, Faculty of Medicine, Hebrew University, Jerusalem, Israel.
Maray
T
T
Davidson
I
I
Malkinson
M
M
Becker
Y
Y
D000818:Animals; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003429:Cross Reactions; D004279:DNA, Viral / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D015723:Gene Library; D006564:Herpesviridae / Q000235:genetics*; D008381:Herpesvirus 2, Gallid / Q000235:genetics; D014176:Protein Biosynthesis; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D014158:Transcription, Genetic; D014422:Turkeys / Q000382:microbiology; D014764:Viral Proteins / Q000235:genetics*
D004279:DNA, Viral; D014764:Viral Proteins; D012313:RNA