期刊文献+

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

Preliminary Application of Quantitative Real-time PCR Assay for Detection of Phosphodiesterase 7A mRNA among Various Porcine Tissues
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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。 The standard curves of recombinant plasmid, containing the target gene fragment (PDE7A) were successfully con- structed 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 X 10^2 copies/μL. The coefficient of variation for inter-experimental re- producibility 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.
出处 《北京农学院学报》 2012年第2期35-38,共4页 Journal of Beijing University of Agriculture
基金 北京市属高等学校人才强教计划资助项目(PHR201107134)
关键词 磷酸二酯酶7A 荧光定量PCR 标准曲线 环磷酸腺苷 phosphodiesterase 7A real-time PCR standard curve cAMP
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参考文献14

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