| 马铃薯栽培品种‘合作88’细菌人工染色体文库的构建与评价 |
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| 引用本文： | 杨煜,杨晓慧,李灿辉,郭晓,单伟伟,马伟清,黄三文,李广存.马铃薯栽培品种‘合作88’细菌人工染色体文库的构建与评价[J].园艺学报,2015,42(2):361-366. |
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| 作者姓名： | 杨煜 杨晓慧 李灿辉 郭晓 单伟伟 马伟清 黄三文 李广存 |
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| 作者单位： | 1山东省农业科学院蔬菜花卉研究所，山东省设施蔬菜分子生物学重点实验室，济南 250100；2云南师范大学生命科学学院，昆明 650500；3中国农业科学院蔬菜花卉研究所，北京 100081 |
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| 基金项目： | 国家‘863’计划项目(2013AA102603-4);国家科技支撑计划项目(2012BAD02B05-10);山东省良种工程农业生物资源创新利用项目(PTBR2013);公益性行业(农业)科研专项(201303018-03);山东省农业科学院青年科研基金项目(2014QNZ03) |
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| 摘 要： | 以四倍体马铃薯栽培品种‘合作88’培养12 ~ 15 d苗龄的幼嫩叶片为试材，提取高分子量基因组DNA，利用限制性内切酶HindⅢ部分酶切后回收100 ~ 300 kb片段，连接到载体CopyControlTM pCC1BACTM的HindⅢ位点上，构建了细菌人工染色体（BAC）文库。该文库约由850 000个克隆组成，空载率约2.08%，保存在1 700个混合池中，克隆插入片段平均大小约为90 kb，覆盖单倍型马铃薯基因组约85倍。随机挑取6个BAC克隆连续培养5 d，提取DNA进行HindⅢ完全酶切检测，确认不同培养时间的BAC克隆的指纹图谱完全一致，表明文库较好的稳定性。BAC文库的构建为马铃薯重要农艺性状基因克隆、基因组测序以及比较基因组研究等奠定了基础。
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| 关 键 词： | 马铃薯 细菌人工染色体文库 抗病性 |
Construction and Characterization of a Bacterial Artificial Chromosome Library of Potato Cultivar C88 |
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| YANG Yu,YANG Xiao-hui,LI Can-hui,GUO Xiao,SHAN Wei-wei,MA Wei-qing,HUANG San-wen,LI Guang-cun,.Construction and Characterization of a Bacterial Artificial Chromosome Library of Potato Cultivar C88[J].Acta Horticulturae Sinica,2015,42(2):361-366. |
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| Authors: | YANG Yu YANG Xiao-hui LI Can-hui GUO Xiao SHAN Wei-wei MA Wei-qing HUANG San-wen LI Guang-cun |
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| Institution: | 1.Institute of Vegetables and Flowers，Shandong Academy of Agricultural Sciences，Molecular Biology Key Laboratory of Shandong Facility Vegetable，Ji’nan 250100，China；2College of Life-Sciences，Yunnan Normal University，Kunming 650500，China；3Institute of Vegetables and Flowers，Chinese Academy of Agricultural Sciences，Beijing 100081，China |
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| Abstract: | A bacterial artificial chromosome library of potato（Solanum tuberosum L.）was constructed for‘C88’，a tetraploid potato cultivar conferring resistance to late blight，PVX and PVY，using BAC vector CopyControlTM pCC1. Fresh leaf tissues from 12–15 day-old seedlings were used to isolate high molecular weight genomic DNA，and 100–300 kb partially HindⅢ digested DNA fragments were selected and ligated into the HindⅢ site of pCC1. The library consists of 850 000 clones and stored in
1 700 pools. The average insert size of BAC clones is about 90 kb and the empty clones consist of about
2.08%. The library is equivalent to 85 haploid potato genomes. Six randomly picked clones showed stable HindⅢ fingerprints after 5 days’ successive culture，indicating that the clones in the library are stable. The library will facilitate gene cloning，sequencing and comparative genomics research of potato. |
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| Keywords: | potato bacterial artificial chromosome library pathogen resistance |
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