猪伪狂犬病病毒XIN-W株gD和gE基因的克隆及序列分析

Clone and Sequencing of gD and gE Gene of PRV XIN-W Isolate of Pseudorabies virus

根据已发表的PRVgD、gE基因序列 ,设计合成了两对引物 ,对猪伪狂犬病病毒XIN W株相关的毒力基因gD、gE序列进行测定和分析。经PCR扩增分别得到了全长为 12 0 9bp,1 743bp的gD、gE全基因序列 ,将它们克隆到pGEM T Easy载体中 ,并转化JM1 0 9,挑取阳性菌落的质粒进行PCR、酶切、测序鉴定 ,与其他参考毒株的gD、gE基因进行比较。结果表明 ,XIN W株与其它毒株相比 ,gD、gE基因核苷酸序列的同源性分别为 97.9%～99.3 % ,98.0 %～ 98.7% ;氨基酸序列的同源性分别为 97.0 %～ 99.3 % ,97.1 %～ 98.1 %。不同的PRV毒株间gD、gE基因在核苷酸水平和氨基酸水平高度保守。遗传进化树显示 ,XIN W株和国内其它毒株起源相同 。

According to the published PRV gD and gE gene nucleotide sequence,two pairs of specific primers were designed and synthesized. A 1 209 bp gD and a 1 743 bp gE gene completed sequence were amplified by polymerase chain reaction (PCR) and colned into pGEM-T-Easy vector. After transforming the recombinant plasmid to JM109,positive colnes were selected and confirmed by PCR and restrict enzyme identification.The sequences of nucleotide and predicted amino acid were compared with those of other published PRV strains.The results showed that gD and gE gene nucleotide sequence homology were 97.9%-99.3%,98.0%-98.7%;the amino acid sequence homology were 97.0%-99.3%,97.1%-98.1%.At the nucleotide and amino acid level, gD and gE gene sequences were respectively compared with other published PRV strains, their homology were very high. Molecular phylogenetic tree showed that XIN-W and other PRV strains in China might originate from the same source and belong to the same evolution embranchment.