CAJ | 학술논문

悬铃叶苎麻（Boehmeria tricuspis （Hance）Makino）是一种多年生草本植物，含有较多的多酚、多糖等次生代谢物质，这些次生代谢物质能够与基因组 DNA 结合从而影响其 DNA 的提取。本文以悬铃叶苎麻幼嫩叶片为试材，采用6种方法提取基因组 DNA，并比较其提取效果，以期寻找到正确的、快速的、能够得到高质量的基因组 DNA 的方法。琼脂糖凝胶电泳、紫外分光光度仪、SSR －PCR 扩增和限制性内切酶酶切等方法检测不同方法提取的全基因组 DNA 的质量和纯度。结果显示：采用方法1（TianGen 试剂盒）提取的溶液中含有杂质，DNA 含量较少；方法2（常规CTAB 法）提取的 DNA 含有较多杂质；方法3（硅胶－CTAB 法）和方法4（酚抽提－CTAB 法）没有提取到 DNA；方法5（高盐预先去杂－CTAB 法）得到的基因组 DNA 溶液含有较多的蛋白质或盐；方法6（葡萄糖预先去杂－CTAB 法）是唯一能获得高质量的基因组 DNA，能够满足 PCR、限制性内切酶酶切等分子实验对 DNA 的要求。本研究找到了一种通用、简单易行，成本低且安全的去除植物 DNA 提取过程中多糖等杂质的方法，为悬铃叶苎麻后续的其他分子生物学的研究奠定了基础。
현령협저마（Boehmeria tricuspis （Hance）Makino）시일충다년생초본식물，함유교다적다분、다당등차생대사물질，저사차생대사물질능구여기인조 DNA 결합종이영향기 DNA 적제취。본문이현령협저마유눈협편위시재，채용6충방법제취기인조 DNA，병비교기제취효과，이기심조도정학적、쾌속적、능구득도고질량적기인조 DNA 적방법。경지당응효전영、자외분광광도의、SSR －PCR 확증화한제성내절매매절등방법검측불동방법제취적전기인조 DNA 적질량화순도。결과현시：채용방법1（TianGen 시제합）제취적용액중함유잡질，DNA 함량교소；방법2（상규CTAB 법）제취적 DNA 함유교다잡질；방법3（규효－CTAB 법）화방법4（분추제－CTAB 법）몰유제취도 DNA；방법5（고염예선거잡－CTAB 법）득도적기인조 DNA 용액함유교다적단백질혹염；방법6（포도당예선거잡－CTAB 법）시유일능획득고질량적기인조 DNA，능구만족 PCR、한제성내절매매절등분자실험대 DNA 적요구。본연구조도료일충통용、간단역행，성본저차안전적거제식물 DNA 제취과정중다당등잡질적방법，위현령협저마후속적기타분자생물학적연구전정료기출。
Boehmeria tricuspis (Hance)Makino is a perennial herbaceous plant which contains ex-ceptionally high amounts of polyphenols,polysaccharides,and other secondary metabolites that interfere with DNA isolation.Six methods were compared for the extraction of genomic DNA from young leaves of Boehmeria tricuspis,and we expected to find a correct method which can make us quickly obtained high quality genomic DNA.The total DNAs extracted by different methods were detected by agarose gel elec-trophoresis,the ultraviolet absorbency,the amplification of DNAs with SSR primer and restriction endo-nuclease reaction.The results indicate the solution of genomic DNA extracted by method 1 (Tian -Gen kit)contains impurities and little DNA,method 2(simple CTAB)produces a lot of impurities,method 3 (silica gel -CTAB)and method 4(phenol extracting -CTAB)can’t extract genomic DNAs,and lots of proteins or salts in the solution of genomic DNA is obtained by method 5 (high -salt predeimpurity -CTAB),while the method 6(glucose predeimpurity -CTAB)is the best way to obtain high quality ge-nomic DNA and can meet the requirement for PCR,restriction endonuclease reaction and other DNA mo-lecular experiments.This study found a generally simple,low cost and safe way to remove impurities such as polysaccharides in plant DNA extracted process,which laid the foundation of follow -up study of ram-ie in other molecular biology.