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Partial transcription map of Marek's disease herpesvirus in lytically infected cells and lymphoblastoid cell lines
Author(s) -
Schat K. A.,
Buckmaster A.,
Ross L. J. N.
Publication year - 1989
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910440119
Subject(s) - biology , virology , microbiology and biotechnology , transcription (linguistics) , lytic cycle , cell culture , gene , marek's disease , dna , rna , northern blot , southern blot , genome , provirus , virus , genetics , philosophy , linguistics
Marek's disease herpesvirus (MDV) can cause either a productive‐restrictive or lytic infection, a latent infection or can transform thymus‐derived lymphocytes. RNA was extracted from infected chicken embryo fibroblasts (CEF) or from lym‐phoblastoid tumour cell lines. Some of the infected CEF were treated with 200 μg/ml cycloheximide to identify immediate early (IE) transcripts, and others with I μM l‐(2‐fluoro‐2‐deoxy‐B‐D‐arabinofuranosyl)‐5‐methyluracil (FMAU), an inhibitor of herpesvirus DNA synthesis to identify early transcripts. An extensive Northern blot analysis was carried out using DNA probes spanning almost the complete MDV genome. In the lytically infected CEF at least 66 discrete transcripts were detected, ranging in size from 9.1 kb to 0.6 kb. Eleven IE transcripts were identified, of which 8 were mapped in the genome segment consisting of the IR L , IR s , U s and TR s . Six transcripts were identified as early genes. In the MD lym‐phoblastoid cell lines MDCC‐HPI, a non‐producer cell line, and MDCC‐CU4I, a non‐expression cell line, 4 and 7 transcripts were detected, respectively. These RNAs were transcribed from IE genes located mainly in the repeat sequences flanking U L and U s and in U s . Treatment of the lymphoblas‐toid cell lines with 20 μ.g/ml 5‐iodo‐2‐deoxyuridine resulted in the additional transcription of I RNA species in HP I and 9 in CU41. Most of the transcripts present in lytically infected cells were also detected in MDCC‐CU36, a cell line with a high percentage of antigen‐positive cells (expression cell line).