减蛋综合征病毒NE4株全基因组序列测定与分析

根据减蛋综合征病毒(EDSV-76)AV-127株的全基因序列(序列号BK000404),用Premier5.0软件设计22对引物,利用PCR方法对EDSV-76病毒NE4株的全基因组进行了分段扩增、克隆和序列测定,并用DNAStar分析软件对各片段的测序结果进行拼接,将该序列与GenBank中已登录的相应序列进行同源性分析,并用六邻体蛋白构建进化树.结果表明:NE4株基因组序列全长33214bp,GC含量为43%,与国际标准株AV-127相比,核苷酸序列同源性为99.6%.碱基的插入或缺失主要在非编码区,在100K蛋白编码区距羧基端1/10处插入了1个碱基C,其ORF编码696个氨基酸,而AV-127株100K蛋白编码709个氨基酸,预计其发挥功能的基团在N端.蛋白同源性分析表明,EDSVNE4株主要蛋白与羊腺病毒OAV(Ovine adenovirusD)、牛腺病毒BAV(Bovine adenovi-rusD)和蛇腺病毒SAV(Snake adenovirus)有较高的同源性,而与禽腺病毒Ⅰ群(CELO)的同源性较低,说明NE4株与禽腺病毒Ⅰ群亲缘关系较远,进化树分析也证实了此特性.

Sequencing and analysis on the complete genome of egg drop syndrome virus NE4 strain

According to the published genome sequences of egg drop syndrome virus(EDSV-76)AV127on GenBank(BK000404),twenty-two pairs of primers were designed by software Premier 5.0and the complete genome of egg drop syndrome virus NE4 strain was amplified by PCR.Splicing each fragment by DNAStar software and analyses the homology of EDSV proteins with other adenoviruses registered in GenBank.Hexon protein was used to build the phylogenetic tree.The results showed that the genome sequence of NE4 strain was 33214bp in length and the GC content was 43%.Compared with the genome of international standard strain AV-127,the nucleotide similarity was 99.6%.Insertions or deletions of bases mainly located in the non-coding region.There was a base C inserted in one-tenth from the C-terminal in the 100 Kprotein coding region,the translation was terminated in advance.So the ORF encoding 696 amino acids while the AV-127 strain encoding 709 amino acids.The N terminal was expected to be function region.Protein homology analysis showed that the main proteins have a high homology with OAV(Ovine adenovirus D),BAV(Bovine adenovirus D)and SAV(Snake adenovirus),but low homology with CELO.This indicated that there was a distant genetic relationship between NE4 strain and fowl adenovirusⅠ.Phylogenetic analysis also indicated this feature.