| 摘要: |
| 根据GenBank中的猪抗病毒蛋白PKR、OAS/RNase L及Mx核苷酸序列设计特异性引物,经RT-PCR技术从猪外周血单个核细胞中扩增和克隆了PKR、OAS/RNase L及Mx的101 bp核苷酸片段。测序鉴定后,将重组质粒10倍系列稀释后作为标准模板,通过实时荧光定量PCR方法,建立了抗病毒蛋白PKR、OAS/RNase L及Mx标准曲线及其直线回归方程;该方法具有线性关系好,特异性、敏感性、重复性高等特点;建立的荧光定量PCR方法可检测抗病毒蛋白PKR、OAS/RNase L及Mx mRNA水平。为猪体内抗病毒蛋白PKR、OAS/RNase L及Mx的mRNA水平的定量检测提供必要的技术。 |
| 关键词: 抗病毒蛋白 外周血单个核细胞 实时荧光定量PCR 重组质粒 |
| DOI：10.11841/j.issn.1007-4333.2009.02.025 |
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| 基金项目:国家“973”计划前期研究专项(2007CB116308);;江苏省自然科学基金(BK2008351) |
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| Establishment of standard curves for detection of porcine antiviral protein genes recombinant plasmids using Real-time PCR |
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| Abstract: |
| The current study is to provide powerful tools for the quantification of antiviral proteins(AVP : PKR,OAS/RNase L and Mx) in pigs.Primers were designed and synthesized based on the nucleotide sequences of PKR,OAS/RNase L and Mx genes available in GenBank.101 bp fragments were amplified by RT-PCR from the porcine peripheral blood mononuclear cells(PBMC),and then the fragments were cloned and sequenced.Recombinant plasmids were diluted by 10-fold serial and used as the Real-time PCR standard templates.Their s... |
| Key words: antiviral protein porcine peripheral blood mononuclear cell Real-time PCR recombinant plasmid |
