基于β-AS基因的甘草道地性形成机制研究

Researches on Formation Mechanism of Genuineness of Glycyrrhiza uralensis Based on β-AS Gene

目的:对甘草道地性形成机制进行探索。方法:在GenBank中检索β-香树酯醇合成酶(β-AS)基因,进行序列比对并找出序列保守区,用primer premier 5.0软件设计引物,对3个不同产地甘草材料的β-AS基因进行扩增、测序,用MegAlign软件进行序列分析并构建系统树。在表达差异实验中,对来自3个产地的9份甘草材料进行总RNA提取,逆转录得到cDNA,以甘草18S基因为内参,PCR扩增后进行相对表达量分析。结果:在序列多态性实验中,9个样品的β-AS序列经比对分析共发现108处变异位点,内含子变异位点数高于外显子变异位点数两倍。在表达差异实验中内蒙组、甘肃组和宁夏组的甘草β-AS基因平均相对表达量有显著性差异。结论:不同产地甘草β-AS基因多态性的差异以及表达量的差异,可能是导致甘草道地性形成的原因之一。

Objective：To explore the formation mechanism of genuineness of Glycyrrhiza uralensis.Method：Searching for beta-amyrin synthase（β-AS） gene in Genbank and finding out the conserved region by sequence alignment,then primers were designed by primer premier 5.0 software.β-AS genes of G.uralensis from 3 different origins were amplified and sequenced,which were analysed by MegAlign software.In the differential expression experiment,the total RNA of G.uralensis from 3 different origins were extracted,then cDNA were obtained by reverse transcription.Using 18S gene of G.uralensis as internal reference,relative expression of β-AS genes of G.uralensis from 3 different origins were detected and analysed.Result：The gene polymorphism experiment showed that 108 variable sites were expresent in the nine samples,and the quantity of variable sites of intron was twice than that of exon.The differential expression experiment showed that the average relative expression of β-AS gene of G.uralensis from different origins was significantly different.Conclusion：The differences of gene polymorphism and expression of β-AS gene in G.uralensis from different origins may be one of the important reasons to result in the genuineness of G.uralensis.