论文部分内容阅读
甘草为常用中药材,具有补脾益气、清热解毒、祛痰止咳、缓急止痛和调和诸药等作用。甘草酸是甘草中的主要活性成分,其生物合成途径受到许多酶的调控,其中3-羟基-3-甲基戊二酰CoA还原酶(3-hydroxy-3-methylglutary CoA reductase,HMGR)催化3-羟基-3-甲基戊二酰辅酶A(3-hydroxy-3-methylglutary CoA,HMG-CoA)生成甲羟戊酸(MVA),是该途径中的第一个限速酶。本文克隆了甘草HMGR基因cDNA序列,对其进行序列分析,并与其他物种进行序列比对。构建了原核表达载体,诱导其外源表达及酶促反应,并利用TLC及GC-MS对产物进行检测。结果表明本文克隆得到的HMGR序列能很好的完成特定的酶促反应任务。由于HMGR基因在甘草酸生物合成途径中的重要作用,因此本实验的研究结果对于在分子水平上揭示高品质甘草的形成机制具有一定的意义。
Licorice is commonly used Chinese herbal medicines, with spleen Qi, detoxification, expectorant cough, urgency and pain relief and to reconcile the role of various drugs. Glycyrrhizic acid is the main active ingredient in licorice and its biosynthesis pathway is regulated by many enzymes. Among them, 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) catalyzes 3 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) to produce mevalonate (MVA) is the first rate-limiting enzyme in this pathway. In this paper, we cloned the cDNA sequence of Glycyrrhiza glabra HMGR gene, sequenced it and sequenced it with other species. The prokaryotic expression vector was constructed to induce its exogenous expression and enzymatic reaction, and the product was detected by TLC and GC-MS. The results showed that the HMGR sequences cloned in this paper can fulfill specific enzymatic reaction tasks well. Because of the important role of HMGR gene in the pathway of glycyrrhizin biosynthesis, the results of this experiment are of great significance to reveal the mechanism of the formation of high-quality licorice at the molecular level.