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首页> 外文期刊>Scientia horticulturae >A deep-coverage BAC library for Ponkan mandarin and its further use in screening ethylene regulated genes.
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A deep-coverage BAC library for Ponkan mandarin and its further use in screening ethylene regulated genes.

机译：用于Ponkan普通话的深层BAC库及其在筛选乙烯调控基因中的进一步用途。

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摘要

A deep-coverage bacterial artificial chromosome (BAC) library of Yanxiwanlu Ponkan, a late maturing cultivar, was constructed and evaluated. Results showed that the library contained a total of 61,000 clones. Restriction analysis indicated that the DNA insert sizes ranged from 30 to 170 kb with an average of ~82 kb. No empty clone was found. Therefore, the library should cover ~13.6-fold of the citrus genome. Continuous sub-cultivation of 5 randomly chosen clones for up to 100 generations showed no detectable change in their restriction profiles. To demonstrate the application of the library in discovery of genes, 7 ethylene-regulated genes were screened by PCR amplification from hierarchically pooled library clones with gene-specific primers. Five positive clones, each representing a different gene, were thus identified from 3840 clones. Sequencing of the clones showed that all of them matched the expected genes, indicating that the library was highly representative.
机译：构建并评估了晚熟品种烟溪万路蓬坎的深层细菌人工染色体（BAC）文库。结果显示该文库包含总共61,000个克隆。限制性酶切分析表明，DNA插入片段大小在30到170 kb之间，平均约为82 kb。找不到空克隆。因此，该文库应覆盖柑橘基因组的〜13.6倍。对5个随机选择的克隆进行连续亚培养达100代，其限制性酶切图谱未检测到变化。为了证明该文库在发现基因中的应用，通过PCR扩增从具有基因特异性引物的分层合并文库克隆中筛选了7个乙烯调节的基因。因此从3840个克隆中鉴定出五个阳性克隆，每个代表不同的基因。克隆的测序表明它们都与预期的基因匹配，表明该文库具有很高的代表性。

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