Preventive Veterinary Medicine, Northeast Agricultural University, Harbin, PR China.
Intervirology. 2010;53(3):141-5. doi: 10.1159/000274974. Epub 2010 Jan 13.
Duck enteritis virus (DEV) is a member of the family Herpesviridae, the genome of which is not completely analyzed. In order to provide further insights into the genomic organization, we decided to amplify the unknown sequence in the US region since three key glycoproteins are coded in this region. Glycoprotein D encoded by US6 gene is essential for virus entry and plays an important role in protecting animals from lethal challenge. Glycoprotein I and E encoded by US7 and US8 gene are required for cell-to-cell spread.
Single oligonucleotide nested polymerase chain reaction was adopted to amplify the unknown sequence. The open reading frames (ORF) were predicted by Editseq program. Then the sequence was submitted to an online program to search promoter, polyadenylation signal and potential N-glycosylation sites. The phylogenetic analysis was performed by MEGA4.1, while multiple alignments were performed by ClustalX.
Three complete ORFs including US6, US7 and US8 were determined. The three genes shared the same polyadenylation signal. Phylogenetic analysis showed that DEV was closed to mardivirus of alphaherpesvirus.
These results extend our understanding of the DEV genome and its classification, and also provide further basic knowledge for functional investigations on the proteins in the US region.
鸭肠炎病毒 (DEV) 是疱疹病毒科的一员,其基因组尚未完全分析。为了进一步了解基因组组织,我们决定扩增 US 区的未知序列,因为该区域编码了三个关键糖蛋白。US6 基因编码的糖蛋白 D 对于病毒进入至关重要,在保护动物免受致死性挑战方面发挥着重要作用。US7 和 US8 基因编码的糖蛋白 I 和 E 是细胞间传播所必需的。
采用单核苷酸巢式聚合酶链反应扩增未知序列。Editseq 程序预测开放阅读框(ORF)。然后将序列提交在线程序搜索启动子、多聚腺苷酸化信号和潜在的 N-糖基化位点。MEGA4.1 进行系统发育分析,ClustalX 进行多重比对。
确定了三个完整的 ORF,包括 US6、US7 和 US8。这三个基因具有相同的多聚腺苷酸化信号。系统发育分析表明,DEV 与α疱疹病毒的马立克病毒密切相关。
这些结果扩展了我们对 DEV 基因组及其分类的理解,也为 US 区蛋白的功能研究提供了进一步的基础知识。