伪狂犬病病毒鄂A株TK基因的克隆及其鉴定

合成了 １ 对能对伪狂犬病病毒（ Ｐｓｅｕｄｏｒａｂｉｅｓ ｖｉｒｕｓ， Ｐ Ｒ Ｖ） Ｔ Ｋ（ｔｈｙｍ ｉｄｉｎｅ ｋｉｎａｓｅ）基因＋ １１９～＋ １ ０７１区进行特异扩增的引物，用猪 Ｐ Ｒ Ｖ 鄂 Ａ 株细胞培养物提取的基因组作模板，扩增出 ９５３ ｂｐ 长的片段，用地高辛标记该片段作探针，通过 Ｓｏｕｔｈ ｅｒｎ 杂交，从克隆有 Ｐ Ｒ Ｖ 鄂 Ａ 株的 Ｂａｍ Ｈ Ｉ片段的重组质粒中钓出含 Ｔ Ｋ 基因的重组质粒 ｐ Ｓ Ｔ Ｋ。对 ｐ Ｓ Ｔ Ｋ 进行酶切分析，绘制了含 Ｔ Ｋ 基因的 Ｂａｍ Ｈ Ｉ片段的图谱，通过测序得出了 Ｔ Ｋ 基因的全序列。将该序列与 Ｐ Ｒ Ｖ Ｎ Ｉ Ａ３ 株 Ｔ Ｋ 基因进行比较，发现鄂 Ａ 株的 Ｔ Ｋ 基因存在变异。

Cloning and Identification of the TK of Gene Pseudorabies Virus Ea Strain

A pair of primers,which can specifically amplified the region form 119 1 071 of PRV TK gene was synthesized,then labeling the PCR product by digoxin as probe.The recombinant plasmid,named pSTK,which contained TK gene,was selected from the libraries by southern blotting hybridization.The Bam HI fragment containing TK gene was analysed by digestion and the sequence of TK gene of PRV Ea strain was determined by dideoxygen temination.The result showed that the map of restriction endonuclease of the Bam HI fragment was consistent with the patterns reported.It is found that variation exists in TK gene of PRV Ea strain.