Peng Q, Zeng M, Bhuiyan Z A, Ubukata E, Tanaka A, Nonoyama M, Shirazi Y
Laboratory of Tumor Virology, Salem-Teikyo University.
Virology. 1995 Nov 10;213(2):590-9. doi: 10.1006/viro.1995.0031.
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.
我们使用与马立克氏病病毒(MDV)基因组BamHI-I2片段右端对应的探针,从MKT-1(一种经MDV转化并潜伏感染的淋巴母细胞系)来源的cDNA文库中分离并测序了两个大小分别为2674 bp和677 bp的cDNA。较大的cDNA克隆代表一种丰富的转录本,它从MDV基因组BamHI-I2的右端延伸到相邻的BamHI-Q2和BamHI-L片段,并包含Meq(MDV Eco-Q)开放阅读框。较小的cDNA克隆代表一种剪接转录本,包含Meq的假定DNA结合结构域以及BamHI-L区域的序列。我们制备了针对Meq部分蛋白质序列的多克隆抗体,并在MKT-1细胞和被MDV裂解感染的细胞中检测到一种44 kDa的蛋白质。此外,与每个cDNA特异序列以及cDNA之间共享序列对应的核糖探针,在被MDV裂解或潜伏感染的细胞中检测到了许多转录本。我们的结果表明,Meq转录单元延伸到BamHI-L片段,并且定位到BamHI-I2右端以及相邻的BamHI-Q2和BamHI-L片段的转录本在潜伏期间并非优先表达。