【摘要】 为了研究当归芍药散(danggui shaoyao san)对鸡卵细胞卵黄生成受体基因(OVR)在鸡卵泡中表达量的影响,采用SYBR Green I法建立检测鸡OVR基因表达水平的实时荧光定量PCR方法。根据GenBank数据库中提供的OVR基因mRNA序列,运用Oligo7软件在基因保守区域设计一对引物,以实验室构建的重组质粒为标准品,β-actin为内参基因,分析OVR基因的表达量。结果表明,该方法扩增效率为102%,检测范围Ct值为10~27,熔解曲线显示为特异的单峰,检测范围间有良好的线性关系(R2=0.999 14)。通过荧光定量PCR检测添加1%、1.5%、2%、2.5%、3%5个浓度当归芍药散的鸡卵泡,发现饲料中当归芍药散添加量为2%时,对OVR基因有极显著影响(P<0.01)。当归芍药散可调控蛋鸡的OVR基因表达,影响蛋鸡产蛋率。更多还原

【Abstract】 In order to study the effect of danggui shaoyao san for OVR gene expressed in chicken follicular,the SYBR-Green I quantitative real-time PCR(QPCR) method was established to detect OVR gene mRNA expression levels. A set of primer was designed by Oligo7 according to the chicken OVR mRNA sequence in GenBank, the positive recombinant plasmid was used as quantitative template to generate standard curve,β-actin gene was used for reference gene, OVR gene mRNA expression levels ware analyzed useing this method. The results showed this method amplification efficiency is 102%,Ct value was from 10 to 27,the melting curve show a single peak,a good linear relationship between detection range(R2=0.999 14),useing this method to detect chicken follicular fed with 1%, 1.5%, 2%, 2.5%, 3% levels of danggui shaoyao san, the study found that the addition amount of 2% danggui shaoyao san had a significant effect on OVR gene(P<0.01). Danggui shaoyao san can regulate the OVR gene expression of laying hens,affect the laying rate.更多还原