猪瘟病毒石门株基因组全长cDNA的克隆与序列分析

Complete genome and sequence analysis of classical swine fever virus strain Shimen/HVRI

参照已发表的猪瘟病毒基因组序列设计合成了9对引物,通过RT-PCR从感染猪瘟病毒的猪全血中扩增得到了覆盖猪瘟病毒石门株基因组全长的9个cDNA片段,将所得片段分别克隆至pOK12载体和pMD18-T载体,经测序和拼接后,获得了猪瘟病毒石门株基因组全序列。序列分析表明,猪瘟病毒石门株基因组全长12297个碱基,含有一个大的开放阅读框架,编码1个由3898个氨基酸残基组成的聚合蛋白,其5'非编码区(5'UTR)和3'非编码区3'(UTR)分别由373和227个碱基组成,与已发表的2个猪瘟病毒石门株的全序列相比,核苷酸同源性分别为99.4%和99.6%,氨基酸同源性分别为99.4%和99.7%,在3'末端第12133位T碱基发生缺失。比较了27个猪瘟病毒全序列的3'UTR,结果提示12133位碱基T缺失区域可能是猪瘟病毒的高变异区。

Nine eDNA fragments covering the complete genome of classical swine fever virus （CSFV） Shimen/HVRI strain were cloned and sequenced, and its complete genomic sequence was assembled. The genome of Shimen/HVRI strain consists of 12 297 bp, and contains a large open reading frame encoding a polyprotein of 3 898 amino acids, which is flanked by untranslated regions （UTRs）, 373 bases at the 5＇-end and 227 bases at the 3＇-end. Homology comparison between the sequence of Shimen/HVRl strain and thosc of Shimen/WD and cF114 showed 99.4 % and 99.6 % identities, respectively, at nucleotide level, and 99.4 % and 99.7 % identities, respectively, at amino acid level. A T deletion in the 3＇UTR of Shimen/HVR1 sequence was noted. Phylogenetic analysis based on 3＇UTR of complete sequences of 27 CSFVs suggests this site may be vulnerable to variation.