| 荧光定量PCR检测牛性别相关基因DAX1表达的标准质粒和标准曲线的构建 |
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| 引用本文： | 徐超,,杜卫华,王宗礼,余大为,王栋,郝海生,赵学明,朱化彬.荧光定量PCR检测牛性别相关基因DAX1表达的标准质粒和标准曲线的构建[J].中国畜牧兽医,2010,37(9):94-99. |
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| 作者姓名： | 徐超 杜卫华 王宗礼 余大为 王栋 郝海生 赵学明 朱化彬 |
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| 作者单位： | （1.中国农业科学院草原研究所，呼和浩特 010010； 2.中国农业科学院北京畜牧兽医研究所，北京 100193） |
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| 摘 要： | 本研究根据GenBank中登录的剂量敏感的性别反转－先天性肾上腺发育不良基因1(dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1，DAX1)设计引物，构建包含DAX1基因cDNA片段的质粒，作为中国荷斯坦牛DAX1基因mRNA定量检测的标准品，建立了DAX1基因mRNA表达实时荧光定量PCR检测方法。结果表明，该方法特异性好，检测灵敏度达102拷贝，线性范围为102～106拷贝，阈值循环数(Ct)与PCR体系中起始模板量的对数值间有着良好的线性关系(r=0.99975),扩增效率高(E=100%)，可以作为检测牛DAX1基因mRNA定量检测方法。
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| 关 键 词： | 牛生殖嵴 DAX1 荧光定量PCR TaqMan探针 标准曲线 |
Construction of Bovine Sex Related Genes DAX1 Standard Plasmid and Standard Curve by Real-time RT-PCR |
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| XU Chao,,DU Wei-hua,WANG Zong-li,YU Da-wei,WANG Dong,HAO Hai-sheng,ZHAO Xue-ming,ZHU hua-bin.Construction of Bovine Sex Related Genes DAX1 Standard Plasmid and Standard Curve by Real-time RT-PCR[J].China Animal Husbandry & Veterinary Medicine,2010,37(9):94-99. |
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| Authors: | XU Chao DU Wei-hua WANG Zong-li YU Da-wei WANG Dong HAO Hai-sheng ZHAO Xue-ming ZHU hua-bin |
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| Institution: | (1.Grassland Research Institute，Chinese Academy of Agricultural Sciences，Hohhot 010010，China；2.Institute of Animal Sciences，Chinese Academy of Agricultural Sciences，Beijing 100193，China) |
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| Abstract: | The primer was designed based on dosage-sensitive sex reversal adrenal hypoplasia critical region on chromosome X gene 1 （DAX1） according to the sequence which was submitted on GenBank. A recombinant plasmid contained cDNA fragment in DAX1 gene was constructed and used for the standard substance for quantitative detection of mRNA of the DAX1 gene in Holstein cows, a real-time PCR was developed for detection of DAX1 gene. The results revealed that this method had a good specificity, and the sensitivity was up to 102, range from 102 to 106. The cutoff value Ct had a good linear correlation with PCR samples. This method could be detected DAX1 gene in bovine quantitatively. |
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| Keywords: | genital ridge DAX1 real-time quantitative PCR TaqMan probe standard curve |
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