目的 为建立三七药材的快速鉴定方法,依据GenBank中人参属248条ITS2序列和本实验获得的46条三七ITS2序列比对结果筛选出可用于三七药材快速鉴定的分子身份证。方法 实验所获得的三七ITS2序列经过Codoncode Aligner软件拼接,与GenBank中下载的人参属序列一起导入MEGA 6.0软件中,使用MUSCLE方法进行比对,查找三七所特有的序列片段。结果 三七ITS2序列长度为230 bp,所有人参属序列比对后长度为258 bp,依据三七与人参属其他物种的SNP位点,开发了一段三七所特有的分子身份证序列,5′-AACCCATCATTCCCTCGCGGGAGTCGATGCGGAGG-3′,如果未知物种扩增产物中含有此段特异序列,就可以判断该物种是三七,否则鉴定为其他物种。结论 本实验开发了一段三七分子身份证序列,可用于三七药材、粉末的快速鉴定,对于鉴定DNA降解严重的材料的有很大的应用前景。
Abstract
OBJECTIVE To establish a fast identification method of Panax notoginseng, molecular signature which can be used as fast authentication of P. notoginseng were screened according to comparison RESULTS of 248 Panax ITS2 sequences from GenBank and 46P. notoginseng ITS2 sequences from this study. METHODS ITS2 Sequences obtained in this study were assembled using Codoncode Aligner, and then were imported to MEGA 6.0 together with Panax ITS2 sequences from GenBank. The comparison was conducted using MUSCLE method and then unique molecular signature of P. notoginseng was found in this comparison results. RESULTS The length of ITS2 sequences of P. notoginseng was 230 bp, and the aligned ITS2 sequences of Panax was 258 bp. Based on one SNP site of P. notoginseng, one unique molecular signature was developed,5′-AACCCATCAT TCCCTCGCGG GAGTCGATGC GGAGG-3′. If unknown samples contained the signature sequence after amplification, it would be identified to be Panax notoginseng and vice versa. CONCLUSION The molecular signature obtained from this study can be used for fast authentication of varying kinds of P. notoginsengcrude drugs and powders and authentication of mixed samples.
关键词
三七 /
分子身份证 /
ITS2 /
鉴定
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Key words
Panax notoginseng /
molecular signature /
ITS2 /
authentication
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中图分类号:
R284
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