伪狂犬病病毒上海株gE和gI基因的克隆及序列分析

参考Genebank发表的伪犬病病毒（Pseudorabies Virus,PRV）的gI和gE基因序列，自行设计并合成了两对引物，对PRV上海株（PRV－SH）进行PCR扩增，产物经琼脂糖电泳分析，均呈现一条约960bp和1740bp的条带，将其克隆入pGEM-T-easy载体中，进行了序旬测定，将PRV－SH株的gI基因与Rice株gI基因比较发现,核苷酸的同源性为94.7%，氨基酸的同源性为91.3%，证实为gI基因，将PRV－SH gE基因序列与Ea株、Ruce株gE基因序列进行比较，结果显示，该序列与PRV Ea株、Rice株gE基因的同源性分别为98.5%、97.5%；的氨基酸序列与Ea株，Rice株和I型单纯疱疹病毒（HSV－1）17株gE的同源性分别为97.2%、94.8%和15.6%。

Cloning and Sequencing Analysis of gE and gI Gene of Pseudorabies Virus SH Strain

According to the Sequences published by the Genebank,two pairs of primer were designed in order to amplify gE gene and partial gI gene of pseudorabies virus Shanghai strain,respectively.The gE gene and parital gI gene were obtained by polymerase chain reaction(PCR),subsequently cloned into pGEM_T vector.The nucleotide and amino acid sequences of SH_strain gI gene were compared with the corresponding sequences of Rice strain,the nucleotide and amino acid homology of SH strain with Rice was 94.7% and 91.3% respectively.It was tested to be gI gene.Analyses of the gE gene nucleotide sequence and of its deduced amino acid sequence were performed with computer programs.The results revealed that the DNA sequence is 1 734base pairs and has the potential to encode a protein of 577 amino acids.The gE gene of PRV_SH strain shares highly conserved nucleotide with PRV Ea and Rice strains and has homology to gE gene of herpes simplex virus type_1(HSV_1).The homology of the nucleotide sequence of PRV_SH strain gE with PRV Ea,Rice strain was 98.5%,97.5%;and the homology of deduced amino among them and HSV_1 was 97.2%,94.8%,15.6%, respectively.