Characterization of Marek's disease virus BamHI-A-specific cDNA clones obtained from a Marek's disease lymphoblastoid cell line.
A cDNA library was constructed from poly(A)+ RNA fractions obtained from a Marek's disease (MD) lymphoblastoid cell line, MDCC-CU41, in which viral gene expression is very limited. Three independent groups (1, 2, and 3) of MD virus (MDV)-specific clones were obtained, which were mapped in the inverted repeat region of the BamHI-A fragment of the MDV genome. Northern blot analysis showed that probes prepared from these cDNA clones hybridized with several transcripts of different sizes in poly(A)+ RNA of MDCC-CU41, although the amounts of these transcripts were relatively small compared to those in MDV lytically infected cells. Moreover, a small open reading frame, which can encode a 94-amino-acid protein, was identified in the A41 cDNA clone (Group 3). By RNase protection assays, the 1.2-kb Group 3 transcriptional unit has been defined. In indirect immunofluorescent antibody assays, antiserum against the bacterially expressed fusion protein, glutathione S-transferase-A41, reacted specifically with the cytoplasmic regions of MDV (strain RB1B)-infected chick kidney cells. However, MDCC-CU41 did not contain a detectable level of the protein determined by these methods.
Ohashi
K
K
Department of Avian and Aquatic Animal Medicine, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853.
O'Connell
P H
PH
Schat
K A
KA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D015245:Deoxyribonuclease BamHI; D008381:Herpesvirus 2, Gallid / Q000235:genetics*; D008380:Marek Disease / Q000382:microbiology; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000382:microbiology; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000235:genetics
D018076:DNA, Complementary; D004279:DNA, Viral; D014764:Viral Proteins; D015245:Deoxyribonuclease BamHI