中文摘要：

为利用实时荧光定量PCR技术研究中草药羊蹄有效化学成分合成相关基因表达情况,采用针对多糖、多酚植物样品的总RNA提取试剂提取高质量的羊蹄总RNA,随后运用反转录PCR方法克隆了羊蹄甘油醛-3-磷酸脱氢酶（GAPDH）基因的部分c DNA序列,并以该序列为模板设计引物,采用实时荧光定量PCR验证其作为内源参照基因的可靠性。结果表明：获得的核酸序列长564 bp,编码188个氨基酸;推导的氨基酸序列与西伯利亚蓼、拟南芥、小麦GAPDH基因编码的氨基酸序列同源性分别为92%、90%和89%;扩增曲线和熔解曲线显示设计的引物可应用于实时荧光定量PCR扩增。羊蹄GAPDH基因的克隆为利用实时荧光定量PCR技术研究目的基因表达情况奠定了基础。

英文摘要：

In order to study the genes expression level of active chemical components in Rumex japonicus Houtt. by real-time fluorescence quantitative PCR,total RNA with reagent for polysaccharide and polyphenol plant samples was extracted,and partial c DNA sequence of GAPDH gene was amplified through reverse transcription PCR. And then a reliability analysis of this sequence as house keeping gene was done by real-time fluorescence quantitative PCR. The results showed that the acquired sequence was 564 bp long and encoded 188 amino acids. The deduced amino acids sequence of GAPDH gene in Rumex japonicus Houtt. was 92%,90% and 89% homologous to that in Polygonum sibiricum,Arabidopsis thaliana and Triticum aestivum,respectively. Melting curve and amplification curve showed that the acquired sequence was suitable as house keeping gene in real-time fluorescence quantitative PCR. This acquired sequence provided a good background on analyzing expression level of genes in Rumex japonicus Houtt. by real-time fluorescence quantitative PCR.