构建甘蓝型油菜Nsa雄性不育系与NR1恢复系可育后代幼小花蕾的全长cDNA文库

构建甘蓝型油菜Nsa雄性不育系与NR1恢复系可育后代幼小花蕾的全长cDNA文库 Construction of the Full-Length cDNA Library of Young Floral Buds from the Fertile Progenies of Brassica napus Nsa Sterile Line and Restorer Line NR1 闫晓红 YAN Xiaohong1 1 first-author 黄进勇 HUANG Jinyong2 2 李晨 LI Chen1 1 胡琼 HU Qiong1 1 魏文辉 WEI Wenhui1 1 中国农业科学院油料作物研究所/农业部油料作物生物学重点开放实验室 中国农业科学院油料作物研究所/农业部油料作物生物学重点开放实验室 Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences/Key Laboratory of Oil Crop Biology of the Ministry of Agriculture Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences/Key Laboratory of Oil Crop Biology of the Ministry of Agriculture 郑州大学生物工程系 郑州大学生物工程系 Department of Bioengineering,Zhengzhou University Department of Bioengineering,Zhengzhou University 以甘蓝型油菜新型恢复系NR1与不育系杂交后代可育植株幼小花蕾为材料,采用SMART（switching mechanism at 5′end of RNA transcript）技术构建了其全长cDNA文库.原始文库的滴度为5.2×107pfu/mL,重组率达96%.随机挑取21个噬菌斑,利用PCR扩增插入片段,电泳结果显示其长度集中在0.7 2 kb之间,平均大小约为1.1 kb.采用铺平板法扩增文库,扩增后的文库滴度为4.3×1012pfu/mL.结果表明,所构建的文库达到了用于分离目的基因的建库要求,为后续恢复基因等重要功能基因全长cDNA的分离克隆奠定了基础. The full-length cDNA library of young floral buds from the fertile progeny plants between the novel Brassica napus Nsa restorer line-1（NR1） and Nsa sterile line had been constructed through SMART（switching mechanism at 5′end of RNA transcript） technique.The results indicated that the titer of the original cDNA library was about 5.2×107 pfu/mL,and the titer of the amplified library was 4.3×1012 pfu/mL.The recombinant rate of the library was about 96%.Twenty-one clones from the library were chosen randomly for detecting cDNA inserts by PCR,showing that the lengths of the inserted fragments were from 0.7 kb to 2 kb,and the average size of insert fragments was about 1.1 kb.These results indicate that the library has high quality,and it is sufficient for the isolations of target genes including fertility restorer gene in the future studies. 甘蓝型油菜 新型恢复系NR1 可育植株 幼小花蕾 全长cDNA文库 Brassica napus novel restorer line NR1 fertile plants young floral buds full-length cDNA library 10.14188/j.1671-8836.2010.03.017 S565.4 国家自然科学基金资助项目（30671312）;湖北省自然科学基金创新群体项目（2008CDA083） 2010-03-01 2021-04-01 3 56