论文部分内容阅读
目的:利用GC-MS方法分析甘草不同3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)基因型表达蛋白的催化效率,为揭示甘草HMGR多态性在优质甘草药材形成中的作用奠定基础。方法:以从甘草中克隆的4种HMGR基因突变型构建表达载体,转化Escherichia coli BL21,进行诱导表达、检测、纯化及体外酶促反应,采用TLC及GC-MS对反应产物进行定性及定量分析。结果:L/V型突变(-HSL,-HSV)催化活性近似,GA插入型突变(GALLV,GALSV)催化活性近似,但插入型突变的催化活性显著高于前者,是前者的2倍左右。结论:甘草功能基因HMGR基因多态性可能是甘草优质药材形成的分子基础。
OBJECTIVE: To analyze the catalytic efficiency of different glycyrrhizic acid coenzyme A reductase (HMGR) genotypes in Glycyrrhiza uralensis Fisch by GC-MS method. In order to reveal the polymorphism of Glycyrrhiza uralensis HMGR polymorphism The role of foundation. Methods: Four kinds of HMGR gene clones were cloned from Glycyrrhiza uralensis, and transformed into Escherichia coli BL21 for inducing, detecting, purifying and enzymatic reaction in vitro. The reaction products were qualitatively and quantitatively analyzed by TLC and GC-MS . Results: The catalytic activities of L / V mutant (-HSL, -HSV) were similar to those of GALLV and GALSV. However, the catalytic activity of insertion mutation was significantly higher than that of the former, which was about 2 times of the former. Conclusion: The polymorphism of licorice functional gene HMGR gene may be the molecular basis of licorice quality medicinal materials.