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

作　　者：孙胜永[1] 李佳[2] 李格宾[1] 姜代勋[2] 陈武[1] 

机构地区：[1]北京农学院动物科学技术学院,北京102206 [2]兽医学(中医药)北京市重点实验室,北京102206

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)

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.

关 键 词：磷酸二酯酶7A 荧光定量PCR 标准曲线 环磷酸腺苷 

分 类 号：S828[农业科学—畜牧学]