应用rRNA基因间隔区碱基测序对中药(大黄)进行鉴定

Identification of Rheum palmatum L(Dahuang)by Method of measuring internal transcribed spacer regions of rRNA gene

目的:对大黄种子中提取的DNA中rRNA基因内转录间隔区进行PCR扩增,对于PCR扩增,并对PCR扩增产物进行碱基序列测定,以期从分子水平建立对中草药的鉴定标准。方法:常规提取当归、大黄种子DNA,利用合成的特异性引物对其DNA中rRNA基因内转录间隔区进行套式PCR扩增,将扩增产物以四色荧光标记的双脱氧末端终止循环法进行碱基序列测定。结果:经琼脂糖凝胶电泳证实大黄种子中rRNA基因内转录间隔区PRC基因内转录间隔区的完整碱基序列。RRNA基因内转录间隔区的碱基序列可作为分子水平鉴定植物中药材的又一有效途径。

Objective:To amplify PCR of the internal transcribed spacer regions of rRNA gene extracted from the DNA of the seed of Rheum palmatum L (Da huang)and then measure the base sequence of the amplified products of DNA,so as to set up an identification standard of Chinese herbal medicines at the molecular level.Method:To extract DNA of the seed of Rheum palmatum L(Da huang)by conventional method and amplify PCR of the internal transcribed spacer regions of rRNA gene in the extracted DNA by composed peculiar primer,and finally measure the base sequence of the amplified products by the method of stopping the circle of the end of double deoxidation marked with four color fluorescent.Result:It is proved by Ago-Gel electrophoresis that PCR amplified products of the internal transcribed spacer regions of rRNA gene exist and that a complete base sequence of the internal transcribed spacer regions of rRNA gene from the seed of Rheum palmatum L(Da huang)is measured.Conclusion:The base sequence of the internal transcribed spacer regions of rRNA gene can be taken as another effective approach to the identification of Chinese herbal materia medica at molecular level.