中文摘要：

根据GenBank中的猪源抗病毒蛋白Mx1基因（poMx1）序列设计1对特异性引物,从Mx1基因全长片段中扩增和克隆了特异性的115 bp核苷酸片段。测序鉴定正确后将重组质粒进行定量并10倍倍比稀释作为扩增模板,通过SYBR GreenⅠ实时荧光定量PCR方法,建立Mx1的标准曲线及回归方程。结果显示：该标准曲线的回归方程为Y=-2.986 3 lg X＋38.239 3（R2=0.998 5）,灵敏度为1×102μL-1;应用该方法对猪瘟兔化弱毒苗接种猪肾细胞（PK-15）后产生的poMx1 mRNA进行定量分析,发现病毒感染细胞4 h后,poMx1的表达量最高（P〈0.01）,然后随着时间的增加而缓慢减少。结论：建立的SYBRGreenⅠ实时荧光定量PCR方法可以检测poMx1 mRNA的表达水平。

英文摘要：

On the basis of the nucleotide sequences of porcine Mx1 gene（poMx1）available in GenBank,a pair of primers was designed and synthesized to amplify a specific 115 bp nucleotide fragment from Mx1 full-length gene.Then,the SYBR Green Ⅰ real-time PCR assay was established based on 10-fold dilution recombinant plasmid as the amplification template after it was cloned and sequenced.The results showed that the standard curve regression equation was Y=-2.986 3 lg X＋38.239 3（R2=0.998 5）,and that the standard curve was of high linearity,specificity,sensitivity and reproducibility.The production of poMx1 mRNA from virus-infected cells was quantitatively analyzed after swine kidney cells（PK-15）was stimulated by classical swine fever virus（HCLV strain）.The data showed the production of poMx1 mRNA was much more after 4 h（P0.01）,and then slowly reduced as time grew.So the results showed that this PCR assay could be applied to detect the expression level of poMx1 mRNA.