贝母的分子生物学鉴定方法的研究

用ＤＮＡ指纹图谱、ＲＡＰＤ和ＤＮＡ测序等技术鉴定中药材已有不少报道[1～ 5] 。ＤＮＡ的提取和ＰＣＲ扩增是分子生物学最基本的操作步骤之一。但是由于商品药材多数经过加工处理 ,ＤＮＡ可能会受到不同程度的降解 ,再者植物药材中含有的次生代谢产物如多糖很难与ＤＮＡ和ＲＮＡ分开 ,而多糖对酶有抑制作用 ,因此将会影响ＤＮＡ的提取和ＰＣＲ扩增。本文以贝母为研究对象 ,探讨不同ＤＮＡ提取方法、不同扩增条件对ＰＣＲ产物的影响 ,为生药的分子生物学鉴定提供参考。材料与方法1　材料　托里贝母Ｆｒｉｔｉｌｌａｒｉａｔｏｒｔｉｆｏｌ…

MOLECULAR BIOLOGICAL IDENTIFICATION OF THE CHINESE DRUG "BEIMU" (BULBUS FRITILLARIAE)

AIM: In order to identify “Beimu” by molecular biology technology, the extraction method of genomic DNA and PCR amplification were studied. METHODS: The genomic DNA extraction methods by SDS, CTAB and DNA_zol reagent were compared, so were DNAs from samples treated with different methods. The effects of DNA quality, annealing temperature and activity of Taq DNA polymerase on PCR products were studied also. RESULTS: The SDS method is simpler and cheaper. The fragment of genomic DNA extracted from fresh sample was longer and purer. The annealing temperature and activity of Taq DNA polymerase showed little effect on PCR amplification, while the quality of DNA affected the PCR amplification directly. CONCLUSION: By extracting genomic DNA from fresh sample and PCR amplification, a specific band was obtained.