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从一株马立克氏病病毒(MDV)转化的淋巴母细胞系中分离和鉴定MDV cDNA:鉴定与MDV Eco-Q蛋白(Meq)反义的开放阅读框。

Isolation and characterization of Marek's disease virus (MDV) cDNAs from a MDV-transformed lymphoblastoid cell line: identification of an open reading frame antisense to the MDV Eco-Q protein (Meq).

作者信息

Peng Q, Shirazi Y

机构信息

Laboratory of Tumor Virology, Salem-Teikyo University/Tampa Bay Research Institute, St. Petersburg, Florida 33716, USA.

出版信息

Virology. 1996 Jul 15;221(2):368-74. doi: 10.1006/viro.1996.0388.

Abstract

Two Marek's disease virus (MDV) cDNAs of 852 and 1168 bp, which map to the right end of the BamHI-I2 fragment of the MDV genome, were isolated from a cDNA library derived from the MDV transformed lymphoblastoid cell line MKT-1. These cDNAs hybridized to relatively abundant leftward mRNA transcripts in MKT-1 cells and cells lytically infected with MDV. The transcriptional initiation site for these transcripts was located in the adjacent BamHI-Q2 fragment, as determined by RNase protection and primer extension assays. A computer search for the presence of leftward open reading frames (ORFs) revealed two ORFs encoding 135- and 195-amino-acid polypeptides. A polyclonal antibody raised against a protein sequence in the N-terminus of the latter ORF detected a 23-kDa protein in the nuclear fraction of MDV-transformed lymphoblastoid cells. Furthermore, this ORF was antisense to part of the MDV Eco-Q protein (Meq) sequence.

摘要

从源自马立克氏病病毒(MDV)转化的淋巴母细胞系MKT - 1的cDNA文库中分离出两个分别为852 bp和1168 bp的MDV cDNA,它们定位于MDV基因组BamHI - I2片段的右端。这些cDNA与MKT - 1细胞以及被MDV裂解感染的细胞中相对丰富的向左mRNA转录本杂交。通过核糖核酸酶保护和引物延伸试验确定,这些转录本的转录起始位点位于相邻的BamHI - Q2片段中。对向左开放阅读框(ORF)的计算机搜索显示有两个ORF,分别编码135个和195个氨基酸的多肽。针对后一个ORF N端蛋白质序列产生的多克隆抗体在MDV转化的淋巴母细胞的核部分中检测到一种23 kDa的蛋白质。此外,该ORF与MDV Eco - Q蛋白(Meq)序列的一部分呈反义关系。

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