Transcriptional analysis of Marek's disease virus (MDV) genes in MDV-transformed lymphoblastoid cell lines without MDV-activated cells.
Spontaneously activated MDV is rarely included in MDV-transformed cells, while it may influence the results of transcriptional analysis. A population consisting of 10(3) MDV-transformed cells probably did not include spontaneously activated MDV, since the estimated frequency of MDV-transformed cells including activated MDV was below 0.01% according to limiting-dilution polymerase chain reaction (PCR) and the presence of the major early antigen pp38 in 6 transformed cell lines. Reverse transcriptase-PCR (RT-PCR) products corresponding to ICP27, pol, TK, US3, A41, gA, gB and UL50 genes were undetectable in 10(3) cells by Southern hybridization of the RT-PCR products. Transcripts of the VP16 and SORF2 genes were detected in the 10(3) cells of MSB-1, and the pp14 gene transcript was found in 10(3) cells of RPL-1 but not in 10(3) cells of HPRS-1, MOGA-2, MSB-1 or MTB-1. A transcript corresponding to the ICP4 sequence was detected as a 0.7 kbp RT-PCR product in 10(3) cells of these MDV cell lines but not in the retrovirus-transformed 1104B1 cell line. The transcript corresponding to the 0.7 kbp RT-PCR product suggested a splice by its size and sequence. Thus, transcriptional analysis of 10(3) MDV-transformed cells revealed that the transcript corresponding to the ICP4 sequence was a common transcript in latently infected MDV-transformed cells, while most of the genes did not transcribe in these cells.
Laboratory of Radiation Biology, Hokkaido University, Sapporo, Japan.
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Ui
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Endoh
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Cho
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Kon
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Iwata
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Maki
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Sato
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Kuwabara
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017931:DNA Primers; D005814:Genes, Viral; D008381:Herpesvirus 2, Gallid / Q000235:genetics*; D008214:Lymphocytes; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D015534:Trans-Activators / Q000235:genetics; D014158:Transcription, Genetic; D014764:Viral Proteins
D017931:DNA Primers; D009687:Nuclear Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C075882:ICP4 protein, Gallid herpesvirus 2