| 猪白细胞介素-8荧光定量PCR检测方法的建立 |
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| 引用本文： | 朱荣昌,林树伯,李俊奇,孙雅红,王月江,王志军.猪白细胞介素-8荧光定量PCR检测方法的建立[J].北京农学院学报,2010,25(4):74-75,80. |
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| 作者姓名： | 朱荣昌 林树伯 李俊奇 孙雅红 王月江 王志军 |
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| 作者单位： | 天津市宁河原种猪场,天津,301504;天津市宁河原种猪场,天津,301504;天津市宁河原种猪场,天津,301504;天津市宁河原种猪场,天津,301504;天津市宁河原种猪场,天津,301504;天津市宁河原种猪场,天津,301504 |
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| 摘 要： | 用RT-PCR技术从猪肺泡巨噬细胞总RNA中扩增出猪白介素（IL）-8基因的278 bp cDNA片段,并克隆到pGEM-T Easy载体中,构建成含有猪IL-8 cDNA片段的重组质粒。通过质粒的Real-time PCR方法,建立猪IL-8 cD-NA的Real-time PCR标准曲线。该方法特异性强、灵敏度高、重复性强,可用于猪IL-8 mRNA含量的定量检测。
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| 关 键 词： | 猪 IL-8 荧光定量PCR |
Development of a Fluorescent Quantitative PCR Assay of Porcine Interlukin-8 |
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| ZHU Rong-chang,LIN Shu-bo,LI Jun-qi,SUN Ya-hong,WANG Yue-jiang,WANG Zhi-jun.Development of a Fluorescent Quantitative PCR Assay of Porcine Interlukin-8[J].Journal of Beijing Agricultural College,2010,25(4):74-75,80. |
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| Authors: | ZHU Rong-chang LIN Shu-bo LI Jun-qi SUN Ya-hong WANG Yue-jiang WANG Zhi-jun |
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| Institution: | ZHU Rong-chang,LIN Shu-bo,LI Jun-qi,SUN Ya-hong,WANG Yue-jiang,WANG Zhi-jun (Tianjin Ninghe Primary Pig Breeding Farm,Tianjin 301504,China) |
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| Abstract: | A cDNA segment of 278 base pair(bp) for porcine interleukin(IL)-8 gene was amplified by RT-PCR method from the total mRNA of porcine alveolar macrophages,and was cloned into the pGEM-T Easy vector.Then,one recombinant plasmid with the 278 bp cDNA segment was obtained.After a real-time fluorescent quantitative PCR with the recombinant plasmid,the standard curve of porcine IL-8 cDNA level was constructed.The results showed that the standard curve was of high specificity,sensitivity and reproducibility,and provided a method to quantify porcine IL-8 mRNA. |
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| Keywords: | porcine IL-8 real-time PCR |
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