| 猪Th1/Th2型细胞因子重组质粒实时荧光定量PCR标准曲线的建立 |
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| 引用本文： | 温立斌,何孔旺,杨汉春,郭容利,李成仁,卢维彩,贾付从.猪Th1/Th2型细胞因子重组质粒实时荧光定量PCR标准曲线的建立[J].华北农学报,2009,24(Z1). |
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| 作者姓名： | 温立斌 何孔旺 杨汉春 郭容利 李成仁 卢维彩 贾付从 |
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| 作者单位： | 1. 江苏省农业科学院兽医研究所,江苏,南京,210014
2. 中国农业大学动物医学院,北京,100094 |
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| 基金项目： | 国家"973"计划前期研究专项，江苏省自然科学基金 |
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| 摘 要： | 依据GenBank登录的猪Th1型细胞因子(IL-2、IL-2、IFN-γ)、Th2型细胞因子(IL-4、IL-6、IL-10)的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上述因子的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,建立了它们各自的标准曲线及直线回归方程.结果表明,该方法具有线性关系好,特异性、敏感性、重复性高等特点,为猪体内IL-2、IL-12、IFN-γ、IL-4、IL-6和IL-10的mRNA水平的定量检测,提供了必要的技术手段.
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| 关 键 词： | Th1/Th2型细胞因子 外周血单个核细胞 重组质粒 实时荧光定量PCR |
Establishment of Standard Curves for Detection of Porcine Th1/Th2 Cytokines Genes Recombinant Plasmids Using Real-time PCR |
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| WEN Li-bin,HE Kong-wang,YANG Han-chun,GUO Rong-li,LI Cheng-ren,LU Wei-cai,JIA Fu-cong.Establishment of Standard Curves for Detection of Porcine Th1/Th2 Cytokines Genes Recombinant Plasmids Using Real-time PCR[J].Acta Agriculturae Boreali-Sinica,2009,24(Z1). |
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| Authors: | WEN Li-bin HE Kong-wang YANG Han-chun GUO Rong-li LI Cheng-ren LU Wei-cai JIA Fu-cong |
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| Institution: | WEN Li-bin1,HE Kong-wang1,YANG Han-chun2,GUO Rong-li1,LI Cheng-ren1,LU Wei-cai1,JIA Fu-cong1(1.Veterinary Research Insititute,Jiangsu Academy of Agriculture Science,Nanjing 210014,China,2.College of Veterinary Medicine,China Agricultural University,Beijing 100094,China) |
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| Abstract: | The primers were designed and synthesized based on the nucleotide sequences of porcine Th1/Th2 cytokines available in GenBank.The 101 bp fragments were amplified by RT-PCR from the porcine peripheral blood mononuclear cells(PBMC),and then the fragments were cloned and sequenced.The recombinant plasmids were diluted by 10-fold serial and used as the real-time PCR standard templates.Their standard curves and the corresponding linear regression equations were obtained.The results indicated that the standard cu... |
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| Keywords: | Th1/Th2 cytokines PBMC Recombinant plasmid Real-time PCR |
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