应用大量生物信息学分析工具及RNA斑点杂交技术对本实验室构建鸭瘟病毒(Duck plague virus,DPV)CHv株基团文库时新发现的dUTPase基因(GenBank(登录号DQ486149)进行了分子特性及转录时相分析。研究表明:该基因大小为1344bp,编码447个氨基酸,包含dUTPase家族蛋白的5个保守motifs,排列形式3-1-2-4—5具备Ⅱ型dUTPELse的典型特征。DPV CHv dUTPase基因在25个疱疹病毒参考株之间高度保守,并与α疱疹病毒的氨基酸同源性较之与β、γ疱疹病毒要高。DPV CHv dUTPase基因转录检测发现DPV感染宿主细胞后最早30min可检测到该基因转录产物,随后转录产物量迅速放大.4h达高峰,24h后下降至相对低的水平。该研究为阐明病毒基因表达调控机理提供了有用的实验数据,并对病毒其他基因的发现和研究具有指导性意义。
A large number of bioinformatics analysis tools and the RNA dot-blot the features of the DPV CHv dUTPase gene (GenBank accession number hybridization technique DQ486149), which was were used to identify first discovered when constructing the duck plauge virus (DPV) CHv strain library in our laboratory. The study shows that the DPV CHv dUTPase gene consists of 1344 nucleotides encoding for a polypeptide of 447 amino acid residues. Moreover, the dUTPase of DPV CHv has five conserved motifs as that among dUTPase family proteins, and the arrangement according to 3-1-2-4-5 belongs to the Class Ⅱ dUTPase subfamily. The sequence comparison of dUTPase among DPV CHv and 25 other reference herpesvirus strains show that DPV CHv is more homologous to alphaberpesviruses than to betaherpesviruses or gammaherpesvimses. The transcription analysis of DPV CHv dUTPase gene shows that the transcripts can be firstly detected at 30 minutes after infection, and then the accumulation reaches a peak in 4 hours. Twenty-four hours later, the transcripts descend significantly to a relatively low level. In a word, these experimental results provide valuable experimental data and certain significance for clarifying the expression regulation of virus genes.