利用两个 BAC 文库对甜瓜进行全基因组 BAC 末端测序。
Genome-wide BAC-end sequencing of Cucumis melo using two BAC libraries.
机构信息
Molecular Genetics Department, Center for Research in Agricultural Genomics CRAG (CSIC-IRTA-UAB), Jordi Girona, 18-26, 08034 Barcelona, Spain.
出版信息
BMC Genomics. 2010 Nov 5;11:618. doi: 10.1186/1471-2164-11-618.
BACKGROUND
Although melon (Cucumis melo L.) is an economically important fruit crop, no genome-wide sequence information is openly available at the current time. We therefore sequenced BAC-ends representing a total of 33,024 clones, half of them from a previously described melon BAC library generated with restriction endonucleases and the remainder from a new random-shear BAC library.
RESULTS
We generated a total of 47,140 high-quality BAC-end sequences (BES), 91.7% of which were paired-BES. Both libraries were assembled independently and then cross-assembled to obtain a final set of 33,372 non-redundant, high-quality sequences. These were grouped into 6,411 contigs (4.5 Mb) and 26,961 non-assembled BES (14.4 Mb), representing ~4.2% of the melon genome. The sequences were used to screen genomic databases, identifying 7,198 simple sequence repeats (corresponding to one microsatellite every 2.6 kb) and 2,484 additional repeats of which 95.9% represented transposable elements. The sequences were also used to screen expressed sequence tag (EST) databases, revealing 11,372 BES that were homologous to ESTs. This suggests that ~30% of the melon genome consists of coding DNA. We observed regions of microsynteny between melon paired-BES and six other dicotyledonous plant genomes.
CONCLUSION
The analysis of nearly 50,000 BES from two complementary genomic libraries covered ~4.2% of the melon genome, providing insight into properties such as microsatellite and transposable element distribution, and the percentage of coding DNA. The observed synteny between melon paired-BES and six other plant genomes showed that useful comparative genomic data can be derived through large scale BAC-end sequencing by anchoring a small proportion of the melon genome to other sequenced genomes.
背景
尽管甜瓜(Cucumis melo L.)是一种经济上重要的水果作物,但目前没有公开的全基因组序列信息。因此,我们对代表总共 33024 个克隆的 BAC 末端进行了测序,其中一半来自以前用内切酶生成的甜瓜 BAC 文库,另一半来自新的随机剪切 BAC 文库。
结果
我们总共生成了 47140 个高质量的 BAC 末端序列(BES),其中 91.7%是配对 BES。两个文库都是独立组装的,然后进行交叉组装,得到了最终的 33372 个非冗余、高质量的序列。这些序列被分为 6411 个重叠群(4.5 Mb)和 26961 个未组装的 BES(14.4 Mb),代表甜瓜基因组的约 4.2%。这些序列被用于筛选基因组数据库,鉴定出 7198 个简单序列重复(相当于每 2.6 kb 一个微卫星)和 2484 个其他重复,其中 95.9%代表转座元件。这些序列也被用于筛选表达序列标签(EST)数据库,揭示了 11372 个与 EST 同源的 BES。这表明甜瓜基因组的约 30%由编码 DNA 组成。我们观察到甜瓜配对 BES 与其他六个双子叶植物基因组之间的微同线性区域。
结论
来自两个互补基因组文库的近 50000 个 BES 的分析覆盖了甜瓜基因组的约 4.2%,提供了对微卫星和转座元件分布以及编码 DNA 百分比等特性的深入了解。观察到甜瓜配对 BES 与其他六个植物基因组之间的同线性表明,可以通过大规模 BAC 末端测序来锚定甜瓜基因组的一小部分到其他已测序的基因组上,从而获得有用的比较基因组数据。
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