Gene enrichment in maize with hypomethylated partial restriction (HMPR) libraries
Gene enrichment in maize with hypomethylated partial restriction (HMPR) libraries
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DOI：
10.1101/gr.3362105
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发表时间：
2005-10-01
期刊：
影响因子：
7
通讯作者：
Bennetzen, JL
中科院分区：
文献类型：
--
作者：
Emberton, J;Ma, JX;Bennetzen, JL
开发了一种新技术，以玉米作为测试生物体，协助对任何复杂植物基因组进行基因富集测序。通过使用甲基化敏感限制酶 Hpall (5'-CCGG-3') 和 HpyCH41V (5'-ACGT-3') 进行独立部分消化来构建低甲基化部分限制性 (HMPR) 文库。纯化并克隆 1-4 kb 的片段，然后对来自 10 个独立文库的超过 2000 个克隆进行序列分析。细胞器克隆占每个文库的-10%，但有助于表明没有生成嵌合克隆，并且不同文库中的消化效率为10%-25%。四个独立的 HMPR 文库经过详细分析，显示出非常相似的基因富集和重复缺失程度。与
A new technology was developed to assist gene-enrichment sequencing of any complex plant genome, employing maize as the test organism. Hypomethylated partial restriction (HMPR) libraries were constructed by Using independent partial digestions with methylation-sensitive restriction enzymes Hpall (5'-CCGG-3') and HpyCH41V (5'-ACGT-3'). Fragments of 1-4 kb were purified and cloned, followed by sequence analysis of >2000 clones from 10 separate libraries. Organellar clones comprised -10% of each library but were useful in showing that no chimeric clones were generated and that digestion efficiencies were 10%-25% in different libraries. Four separate HMPR libraries, analyzed in detail, exhibited very similar degrees of gene enrichment and repeat depletion. Known gene homologies were found in similar to 25% of the HMPR clones, compared with