伪狂犬病病毒冀A株TK基因的克隆及序列分析

Cloning and Sequence Analysis of the TK Gene of Pseudorabies virus JA Strain

为了分析比较伪狂犬病病毒(PRV)冀A株TK基因与GenBank中收录的国内外其他PRV毒株TK基因核苷酸及氨基酸序列的同源性,以伪狂犬病病毒冀A株的基因组为模板,PCR扩增了其胸腺激酶(TK)基因,并对其进行了克隆和测序。结果表明,TK基因的开放阅读框(ORF)和所比较的各株的核苷酸和氨基酸同源性都在99%以上。核苷酸序列中发现在起始密码子的上游有3段GC框样的序列,在终止密码子中发现多聚腺苷加尾信号AATAAA;在氨基酸序列中发现有疱疹病毒TK基因的保守序列R*Y*DG**G*GK*T-和-FDRHP*A***C*P*AR-。

The homology of the nucleotide sequence and its deduced amino acid sequence of pseudorabies virus JA strain with other strains which published in GenBank was analyzed. Using the genome DNA of the strain JA of pseudorabies virus as template, the TK gene fragment was amplified by polymerase chain reaction（PCR）. The PCR product was ligated to pUC19 plasmid vector and the nucleotides of the TK gene was sequenced. The result showed that the homology of the nucleotide sequence and amino acid sequence of JA TK ORF with other strains was over 99%. Upstream of the initiation codon, three putative GC boxes are located. A potential poly A signal was detected at downstream from the termination codon. We have identified conserved domains of the thymidine kinase proteins of the alpha herpes viruses： -R ＊ Y ＊ DG＊＊G ＊GK＊T-and-FDRHP＊A＊＊＊C＊P＊AR-.