野生大豆核基因组Mb级DNA的制备与酶切  

Preparing and Digesting Megabase-size Nuclear DNA of Glycine soja Genome

作　　者：李晓玲[1,2] 李克秀[2] 赵洪锟[1] 董英山[1] 

机构地区：[1]吉林省农业科学院生物技术研究中心,吉林长春130033 [2]长春工业大学化学与生命科学学院,吉林长春130012

出　　处：《大豆科学》2011年第1期33-36,40,共5页Soybean Science

基　　金：农业部野生植物资源保护资助项目(2010);农业部CWRC资助项目(CWRC-20080205);国家重点基础研究发展计划"973计划"资助项目(2007CB116205);国家转基因专项资助项目(2008ZX08004-004)

摘　　要：以野生大豆黄化幼苗为材料提取其细胞核DNA,经LMP包埋并用蛋白酶K裂解其中的核蛋白后,采用脉冲电泳回收2 Mb左右细胞核DNA。用HindⅢ对回收细胞核DNA进行部分酶切并用脉冲电泳回收酶切后的DNA片段,经电洗脱、浓缩和透析后DNA溶液浓度可达10 ng.μL-1。连接转化检测结果表明:该DNA可用于后续野生大豆基因组可转化人工染色体(TAC)文库的构建和基因组分析。Nuclear DNA was extracted from etiolated seedlings of Glycine soja.Generally,the nuclei were embeded in low-melting-point agarose pulgs,digested with proteinase K and depurated with pulsed field gel electrophoresis to yield about two-Megabase-size DNA.The concentration of DNA solution could be up to 10 ng·μL-1 after partial digestion by HindⅢ,elution by pulsed field gel electrophoresis,concentration and dialysis.The result of examination by ligation and eletroporation showed that the DNA obtained by the method was suitable for consequent construction of genomic library of Glycine soja.

关 键 词：野生大豆 Mb级DNA 脉冲电泳 基因组可转化人工染色体(TAC)文库 

分 类 号：S565.1[农业科学—作物学]