| 猪磷酸二酯酶7A mRNA荧光定量PCR检测方法的建立与初步应用 |
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| 引用本文： | 孙胜永,李佳,李格宾,姜代勋,陈武. 猪磷酸二酯酶7A mRNA荧光定量PCR检测方法的建立与初步应用[J]. 北京农学院学报, 2012, 27(2): 35-38 |
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| 作者姓名： | 孙胜永 李佳 李格宾 姜代勋 陈武 |
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| 作者单位： | 北京农学院动物科学技术学院,北京,102206;兽医学(中医药)北京市重点实验室,北京,102206 |
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| 基金项目： | 北京市属高等学校人才强教计划资助项目 |
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| 摘 要： | 为研究磷酸二酯酶(PDE)7A基因在猪体内的分布,本试验通过构建含有目的基因片段标准质粒的方法,成功建立荧光定量PCR检测PDE7A基因的标准曲线,可检测到最低1×102copies/μL的初始模板量,重复试验变异系数为2.22%～3.71%,具有较高的灵敏度和可重复性。用该方法检测到PDE7AmRNA在试验用小型猪的心肌中表达量最高(比率为4.54×10-2),骨骼肌次之(1.44×10-2),其他内脏器官较少。
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| 关 键 词： | 磷酸二酯酶7A 荧光定量PCR 标准曲线 环磷酸腺苷 |
Preliminary Application of Quantitative Real-time PCR Assay for Detection of Phosphodiesterase 7A mRNA among Various Porcine Tissues |
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| SUN Sheng-yong , LI Jia , LI Ge-bin , JIANG Dai-xun , CHEN Wu. Preliminary Application of Quantitative Real-time PCR Assay for Detection of Phosphodiesterase 7A mRNA among Various Porcine Tissues[J]. Journal of Beijing Agricultural College, 2012, 27(2): 35-38 |
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| Authors: | SUN Sheng-yong LI Jia LI Ge-bin JIANG Dai-xun CHEN Wu |
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| Affiliation: | 1(1.Department of Animal Science and Technology,Beijing University of Agriculture,Beijing,102206,China;2.Beijing Key Laboratory of Veterinary Medicine,Beijing,102206,China) |
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| Abstract: | The standard curves of recombinant plasmid,containing the target gene fragment(PDE7A) were successfully constructed and utilized in a quantitativeRT-PCR assay for PDE7A expression.qRT-PCR demonstrated high sensitivity with a minimum detection concentration of Initial Template of 1×102copies/μL.The coefficient of variation for inter-experimental reproducibility ranged from 2.22to3.71%.Using qRT-PCR,PDE7A mRNA expression was assessed and compared with various porcine tissues,obtained from Chinese miniature experimental pigs.The results indicated that PDE7A exhibits tissue-specific differences,with highest expression levels in heart and skeletal muscle. |
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| Keywords: | phosphodiesterase 7A real-time PCR standard curve cAMP |
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