伪狂犬病病毒Min-A株TK基因的克隆与序列分析  

Cloning and sequence analysis of the TK gene of pseudorabies virus Min-A strain

作　　者：胡涛[1] 崔保安[1] 王岩[2] 杨明凡[1] 王学斌[1] 张素梅[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]郑州牧业工程高等专科学校,河南郑州450008

出　　处：《河南农业大学学报》2004年第1期111-115,共5页Journal of Henan Agricultural University

摘　　要：参考GeneBank收录的伪狂犬病病毒TK基因的序列设计了1对引物,对PRVMin A株进行了PCR扩增,扩增产物克隆于pGEM TEasy载体.对重组质粒进行限制性内切酶分析和基因测序,证实了克隆片段的可靠性.测序结果表明目的片段包含1个957bp的开放性阅读框(ORF),编码318个氨基酸组成的多肽.在TKORF上游 22,-166,-199位存在3个GC框样序列,在终止密码子下游第110个核苷酸处的1306位存在有多聚腺苷加尾信号.PRVMin A株与PRVEa株、NIA 3株TK的核苷酸同源性分别为98.4%,97.9%;推导氨基酸的同源性分别为97.8%,97.2%.通过对α疱疹病毒TK氨基酸进行多序列比较,获得了在PRVTK氨基酸序列上的6个保守区间.推测这些保守氨基酸对于胸苷激酶(TK)结构的稳定性和催化活性的发挥是必需的.A pair of primers was designed according to the sequences pulished by the GeneBank in order to amplifiy TK gene of the pseudorabies virus Min-A strain.The TK gene was obtained by polymerase chain reaction(PCR), and then cloned into the pGEM-T Easy vector. The recombinant plasmid of pGTE-TK was identified by restriction enzyme analysis and sequencing ,which proved completely its validity. Nucleotide sequencing revealed that this fragment contained an open reading frame of 957 bp encoding a 318 aa protein.Upstream of the TK ORF,three putative GC boxes are located at positions -22,-166 and -199. A potential poly A signal begins 110 nucleotides downstream from the termination codon at position 1 306. The homology of the nucleotide sequence of PRV Min-A strain TK with PRV Ea strain,NIA-3 strain was 98.4%,97.9%,respectively ;and the homology of predicted amino acids among them was 97.8%,97.2%,respectively.We have identified six conserved domains by multiple sequence alignments of the thymidine kinase proteins of the alphaherpesviruses. It is speculated that these conserved amino acid residues are necessary for structural stability or involved in functional domains responsible for catalytic activities.

关 键 词：伪狂犬病毒 胸苷澈酶 克隆 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]