CAJ | 학술논문

为了从富含多酚和多糖的药用植物组织中分离出高质量基因组DNA，本文通过改良几种已经存在的分离方法，获得了一种以CTAB法为基础的分离高质量完整DNA的简便、快速方法。使用该方法从人参、西洋参及三七的新鲜或干燥根组织分离DNA时，A260/A280为1.8左右，分子量大小约为21.2 kb,由此所得的DNA可直接用于AFLP分析，即用EcoRⅠ／MseⅠ消化DNA后,用DNA的限制酶切片段经T4 DNA连接酶连接，再用巢式PCR扩增,构建出可重复的、多态性丰富的人参、西洋参、三七基因组DNA AFLP指纹图谱。结果表明，该技术可有效地从富含多酚和多糖的药用植物组织中分离出高质量和高分子量DNA，此法亦有望用于其它植物DNA的分离。
위료종부함다분화다당적약용식물조직중분리출고질량기인조DNA，본문통과개량궤충이경존재적분리방법，획득료일충이CTAB법위기출적분리고질량완정DNA적간편、쾌속방법。사용해방법종인삼、서양삼급삼칠적신선혹간조근조직분리DNA시，A260/A280위1.8좌우，분자량대소약위21.2 kb,유차소득적DNA가직접용우AFLP분석，즉용EcoRⅠ／MseⅠ소화DNA후,용DNA적한제매절편단경T4 DNA련접매련접，재용소식PCR확증,구건출가중복적、다태성봉부적인삼、서양삼、삼칠기인조DNA AFLP지문도보。결과표명，해기술가유효지종부함다분화다당적약용식물조직중분리출고질량화고분자량DNA，차법역유망용우기타식물DNA적분리。
In order to isolate high-quality genomic DNA from medicinal plant tissues enriching polyphenols and polysaccharides, a simple and rapid method based on CTAB extraction for isolating high-quality intact DNA was established by modifying several existing methods. With this technique, the absorbance ratio (A260/A280) of DNAs obtained from fresh and /or dried roots of Panax ginseng, P. Quinquefolius and P. notoginseng was 1.8 approximately. The restriction fragments of DNAs were directly digested with restriction enzyme (EcoRⅠ/MseⅠ), linked up by T4DNA ligase and amplified by nested PCR. Reproducible amplified fragment length polymorphism (AFLP) genomic DNA fingerprinting profiles were established with the isolated DNAs. The results demonstrate that the modified technique may be efficient and reliable in isolating high-quality and high-molecular-weight DNAs from fresh and/or dried medicinal plants containing a high content of polyphenols and polysaccharides. We expect that this method can also be applied to other plants.