条斑紫菜基因组Fosmid文库构建
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国家“八六三”高技术研究发展计划(2006AA10A402, 2006AA10A413); 国家自然科学基金项目(30700621); 教育部新世纪人才支持计划(NCET060596); 教育部科学研究重点项目(107070);国家自然科技资源共享平台项目(2006DKA30470-016);山东省优秀中青年科学家科研奖励基金(2008BS06002)


Construction of genomic Fosmid library of Porphyra yezoensis
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    摘要:

    高分子量基因组文库是开展条斑紫菜基因组学研究的必备工具。构建了由23 040个Fosmid克隆组成的条斑紫菜孢子体无偏倚基因组文库。检测分析表明:文库克隆重组率为100%;插入DNA片段长度为28~40 kb,平均长度为35 kb,文库覆盖率约为条斑紫菜基因组的2.78倍;从超级池中筛查条斑紫菜18S rRNA、atpA、h2A、rbcL基因,至少能得到一个阳性克隆,印证了计算所推测的文库覆盖率;随机选取6个Fosmid克隆经100代传代后插入DNA片段未发生丢失或长度变化,表明克隆传代的稳定性。该文库的构建为开展条斑紫菜基因组特性分析和基因克隆奠定了基础。

    Abstract:

    Genomic library with highmolecular weight inserts is an essential and powerful tool for genomic studies. An unbiased fosmid library of Porphyra yezoensis was constructed and characterized. The library comprised 23 040 clones with 100% recombination frequency. The range of insert size of most bulk of clones is from 28 kb to 40 kb and the average size is about 35 kb. Therefore, the library coverage is about 2.78 genomeequivalent. The theoretical coverage was verified by successfully screening out 4 randomly selected genes, 18S rRNA, atpA, h2A and rbcL, once at least from the super pools which covered the whole library clones. No variations were observed by comparing the original clones with those of the hundredth generation cultivated successively, indicating the good fidelity and stability of the fosmid library. This study laid the foundations for gene cloning and genomic research in P. yezoensis.

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茅云翔,崔菁菁,孔凡娜.条斑紫菜基因组Fosmid文库构建[J].水产学报,2009,33(3):396~402

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  • 收稿日期:2008-04-27
  • 最后修改日期:2009-01-10
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  • 在线发布日期: 2009-05-07
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