Isolation and characterization of Marek's disease virus (MDV) cDNAs mapping to the BamHI-I2, BamHI-Q2, and BamHI-L fragments of the MDV genome from lymphoblastoid cells transformed and persistently infected with MDV

• Published in: Virology (New York, N.Y.) Vol. 213; no. 2; pp. 590 - 599
• Main Authors: Peng, Q, Zeng, M, Bhuiyan, Z A, Ubukata, E, Tanaka, A, Nonoyama, M, Shirazi, Y
• Format: Journal Article
• Language: English
• Published: United States 10.11.1995
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cell Transformation, Viral; Deoxyribonuclease BamHI; DNA Primers; DNA, Complementary - isolation & purification; DNA, Viral - genetics; DNA, Viral - isolation & purification; Gene Library; Genome, Viral; Herpesvirus 2, Gallid - genetics; Herpesvirus 2, Gallid - physiology; Lymphocytes - virology; Molecular Sequence Data; Restriction Mapping; RNA, Viral - genetics; Transcription, Genetic; Tumor Cells, Cultured; Viral Proteins - analysis; Viral Proteins - genetics; Virus Latency
• ISSN: 0042-6822

We have isolated and sequenced two cDNAs of sizes 2674 and 677 bp from a cDNA library derived from MKT-1, a lymphoblastoid cell line transformed and latently infected with Marek's disease virus (MDV) using probes corresponding to the right-hand end of the BamHI-I2 fragment of the MDV genome. The larger cDNA clone represents an abundant transcript, which extends from the right-hand end of BamHI-I2 to the adjacent BamHI-Q2 and BamHI-L fragments of the MDV genome and contains the Meq (MDV Eco-Q) open reading frame. The smaller cDNA clone represents a spliced transcript containing the putative DNA binding domain of Meq as well as sequences in the BamHI-L region. We prepared a polyclonal antibody against part of the protein sequence of Meq and detected a 44-kDa protein in MKT-1 cells and in cells lytically infected with MDV. In addition, riboprobes corresponding to sequences specific to each cDNA as well as shared sequences between cDNAs detected a number of transcripts in cells either lytically or latently infected with MDV. Our results indicate that the Meq transcriptional unit extends to the BamHI-L fragment and that the transcripts mapping to the right-hand end of the BamHI-I2 and adjacent BamHI-Q2 and BamHI-L fragments are not preferentially expressed during latency.