猪巨细胞病毒福建株DPOL基因的克隆及序列分析

Cloning and Sequencing Analysis of DPOL Gene of Porcine Cytomegalovirus

为了明确猪巨细胞病毒(porcine cytomegalovirus,PCMV)福建株(PCMV-FJ01株)DPOL基因特征,本研究根据GenBank数据库中PCMV DPOL基因序列特征,利用引物设计软件Oligo 7.0设计针对DPOL基因的引物,以PCMV-FJ01株核酸DNA为模板,对其进行分段PCR扩增。将分段扩增的产物经胶回收试剂盒回收后进行克隆测序,对测序结果进行拼接后获得PCMV-FJ01株DPOL基因序列,并进行生物信息学分析。结果表明,PCMV-FJ01株DPOL基因全长为3 021bp,编码1 006个氨基酸;其与GenBank中的PCMV分离株DPOL基因核苷酸和氨基酸同源性分别在98.7%和99.1%以上。遗传进化分析发现,相对于巨细胞病毒属其他种代表株,PCMV分离株DPOL基因在遗传进化上和玫瑰疱病毒属代表株更近。建议根据PCMV DPOL基因遗传进化特征,将其划归为玫瑰疱病毒属的一个病毒新种——猪玫瑰疹病毒(porcine roseolovirus)。

In order to know the DPOLgene characterization of porcine cytomegalovirus（PCMV）（PCMV-FJ01strain）,three pairs of specific primer were designed by Oligo 7.0software based on comparison the characterization of DPOLgene retrieved from GenBank.The target DPOLgene fragments were amplified using PCR methods with the strain PCMV-FJ01 genomic DNA.The target PCR fragments were cloned and sequenced.The assembly sequences were bioinformatics analysis.The DPOL gene of PCMV-FJ01 strain was 3 021 bp in length,coding 1 006 amino acids.The results showed that the homology of the nucleotide sequence and amino acid sequence were above 98.7% and 99.1%,respectively.Phylogenetic analysis revealed that the PCMV was closer with genus Roseolovirus rather than genus Cytomegalovirus.The results suggested that PCMV should be new species under genus Roseolovirus base on the phylogenetic relationship.