伪狂犬病毒粤A毒株TK基因的扩增、克隆与序列测定

Amplification, Cloning and Sequencing of a Fragment of TK Gene of Pseudorabies Virus

以华南农业大学兽医学院传染病教研室分离的伪狂犬病毒粤A毒株 (PRVYA)的基因组为模板 ,利用聚合酶链反应 (PCR)扩增TK基因 ,获得预定大小的片段 ,将这一片段克隆到PMD18-T载体中 .对重组质粒PMD18-TK进行PCR鉴定、限制性内切酶分析和克隆片段的序列测定、比较 ,证实了克隆片段的可靠性 .

Using the genome DNA of the strain YA of pseudorabies virus(PRVYA) as template and with flanking primers, the TK gene fragment was amplified by polymerase chain reaction (PCR). The expected size of the fragment was obtained, and it was then cloned into the PMD18-T vector. The recombinant plasmid PMD18-TK was identified by PCR, restriction enzyme analysis and sequencing, which completely proved its validity.