甘草β-香树酯醇合成酶编码区SNP与甘草酸含量的相关性研究  

Correlation analysis between single nucleotide polymorphism of β-amyrin synthase and content of glycyrrhizic acid in Glycyrrhiza uralensis

作　　者：沈湛云[1] 刘春生[1] 王学勇[1] 呙未[1] 李贝宁[1] 

机构地区：[1]北京中医药大学中药学院,北京100102

出　　处：《中国中药杂志》2010年第7期813-816,共4页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30672615)

摘　　要：目的:探讨甘草β-香树酯醇合成酶(β-amyrin synthase,bAS)编码区SNP与甘草酸含量之间的相关性。方法:采用HPLC法测量80株人工栽培甘草的甘草酸含量,采用SAS 9.0软件将80株甘草按照甘草酸含量极显著水平(P<0.000 1)进行分组;采用RT-PCR技术,扩增出甘草bAS编码区序列,运用DNAman分析软件找出该序列的SNP位点,进而分析该位点与甘草酸含量高低的相关性。结果:bAS基因编码区共有94,254 bp 2个突变位点,在94 bp位点发生G/A转换,为错义突变,导致该位点处甘氨酸/天冬氨酸转换,254 bp处发生C/T转换,为同义突变,根据序列变异将所测样品划分成G-T基因型、A-T基因型、G-C基因型和A/G-C基因型。结论:A-T基因型、G/A-C基因型和G-T基因型和高含量甘草酸形成具有显著的相关性。Objective： To analyze the correlation between content of glycyrrhizic acid and the single nucleotide polymorphism of ？-amyrin synthase （bAS） in Glycyrrhiza uralensis. Method： glycyrrhizic acid content in 80 samples of the cultivated G. uralensis were determined by HPLC; According to the very significant level （P〈0.0001）, 80 samples in accordance with glycyrrhizic acid will be grouped by SAS 9.0; Using RT-PCR strategy to amplification the Open Reading Frame of β-amyrin synthase with the template of total RNA extracted from roots of G. uralensis and then using DNAman to analyze the relationship between glycyrrhizic acid content and the single nucleotide polymorphism of ？-amyrin synthase （bAS）. Result： There exited two mutation sites 94 bp and 254 bp, G/A conversion occurred at 94 bp site, which belonged to a missense mutation. G/A conversion led to the corresponding amino acid conversion （Gly→Asp）; C/T conversion occurred at 254 bp site, which belonged to a synonymous mutation. According to sequence variation, the samples were divided into four genotypes： G-T genotype,A-T genotype,G/A-C genotype and G-T genotype. Conclusion： A-T genotype,G/A-C genotype and G-T genotype are correlated with the high content of glycyrrhizic acid .

关 键 词：甘草酸 β-香树酯醇合成酶 单核苷酸多态性 

分 类 号：R284.1[医药卫生—中药学]