摘要
为了研究当归芍药散(dangguishaoyaosan)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SYBRGreenI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Olig07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—aetin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=0.99914)。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。
In order to study the effect of danggui shaoyao san for OVR gene expressed in chicken fol- licular, 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 mehing curve show a single peak, a good linear relationship between de- tection 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 sha- oyao 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.
出处
《饲料工业》
北大核心
2014年第1期34-37,共4页
Feed Industry