基于PCR-RFLP的西洋参和人参指纹特征鉴定

Fingerprint Dentification of Panax Quinquefolius and Panax ginseng Based on PCR-RFLP

目的建立西洋参与人参限制性片段长度多态性聚合酶链式反应(polymerase chain reaction restriction fragment length polymorphism,PCR-RFLP)的鉴定方法.方法采用改良的CTAB法从西洋参和人参中提取基因组DNA.应用生物信息学软件设计西洋参与人参核糖体DNA ITS序列的特异性引物,利用PCR技术对指定序列进行扩增,并通过RFLP方法酶切后进行指纹图谱的鉴别比较.结果提取西洋参与人参基因组DNA19 kb,并扩增核糖体122 bp大小的基因片段,经酶切后西洋参为80 bp和42 bp长度的两条片段,而人参仍然为122 bp长度的单一片段.结论西洋参与人参DNA具有特异性酶切位点,可作为西洋参与人参鉴定的有效方法,该方法简便快速,结果可靠.

Objective To establish DNA PCR-RFLP method for the identification of Panax quinquefolius and Panax ginseng.Method The modified CTAB method was used to extract genome DNA from Panax quinquefolius and Panax ginseng.Specific primers were designed with bioinformatics software of ribosomal DNA ITS sequences for Panax quinquefolius and Panax ginseng. The specified sequence was amplified by PCR technology,and the enzyme products obtained by the method of RFLP were identified and compared for fingerprint.Results DNA 19 kb genome fragments were obtained from Panax quinquefolius and Panax ginseng. The 122 bp size of the ribosomal gene fragment was amplified. After enzyme digestion,Panax quinquefolius had two fragments of 80 bp and 42 bp,while Panax ginseng had only a single fragment of 122 bp.Conclusion Both the DNA fingerprints of Panax quinquefolius and Panax ginseng had specific enzyme site which could be used as an effective method for identification.The experimental results showed that the method was simple,rapid and reliable.