鸭瘟病毒GD株TK、dUTPase和PK基因的克隆及序列分析

Cloning and Sequencing of TK,dUTPase and PK Genes in Duck Plague Virus GD Strain

根据已发表的鸭瘟病毒(DPV)TK、dUTPase、PK基因序列,设计合成了3对特异性引物,对DPV GD株的3个基因进行PCR扩增,产物克隆至PTA2载体并进行测序。结果表明,TK基因ORF全长为1 077 bp,编码358个氨基酸;dUTPase基因ORF全长为1 344 bp,编码447个氨基酸;PK基因ORF全长为1 158 bp,编码385个氨基酸。与GenBank上其他DPV毒株的同源基因进行比较,TK基因核苷酸的同源性为99.5%～100%,氨基酸同源性为98.9%～100%;dUTPase基因核苷酸的同源性为99.9%,氨基酸同源性为99.8%;PK基因核苷酸的同源性为99.8%～100%,氨基酸同源性为99.7%～100%。这表明TK、dUTPase、PK基因在DPV基因组中高度保守,为构建DPV基因缺失转移载体奠定了基础。

According to the genomic sequences of DPV in GenBank,three pairs of primers were designed for PCR amplication of TK,dUTPase and PK genes in duck plague virus GD strain.The PCR products were cloned into PTA2 vector and sequenced.The result revealed that the ORFs of TK,dUTPase and PK genes were 1 077 bp,1 344 bp and 1 158 bp in size and encoded 358 aa,447aa and 385 aa,respectively.Sequence analysis demonstrated that the nucleotide sequences of TK,dUTPase and PK genes shared 99.5%-100%,99.9% and 99.8%-100% similarities with the published sequences of the other DPV strains,while the amino acid sequence similarities were 98.9%-100%,99.8% and 99.7%-100%,respectively.The result suggested that TK,dUTPase and PK genes of DPV are highly conservative.This study laid the foundation for the construction of transfer vector of duck plague virus.