Nuclear DNA‐based markers for plant evolutionary biology
Nuclear DNA‐based markers for plant evolutionary biology
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DOI：
10.1046/j.1365-294x.1997.00153.x
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发表时间：
1997-02
影响因子：
4.9
通讯作者：
A. Strand;J. Leebens-Mack;B. Milligan
中科院分区：
文献类型：
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作者：
A. Strand;J. Leebens-Mack;B. Milligan
虽然基于DNA的标记可以为植物进化生物学的研究提供丰富的信息，但由于缺乏可用于PCR的引物，进展受到限制。为了克服这一局限性，我们概述了一种用于开发靶向低拷贝数核基因区域的寡核苷酸引物的方案。该方案旨在产生普遍有用的引物组。为了测试我们的方法，我们设计了八个引物组，并测试了它们从每个双子叶植物和一个单子叶植物亚类的代表中扩增靶标的能力。八个引物组中的五个引物组扩增了七个分类群中至少五个分类群的靶标，因此表现出广泛的分类学用途;其余引物相当特异，但是，扩增了最多三个分类群的靶标。仅在一个引物-分类单元组合中产生复杂的多带扩增。总的来说，所概述的方案证明在鉴定靶向低拷贝数核基因的广泛适用的引物方面非常有用。这种方法的广泛应用将有效地大大增加植物核基因组多样性的遗传信息量。
While DNA‐based markers can provide a wealth of information for the study of plant evolutionary biology, progress is limited by the lack of primers available for PCR. To overcome this limitation, we outline a protocol for developing oligonucleotide primers targeting regions of low copy‐number nuclear genes. This protocol is intended to lead to universally useful primer sets. To test our approach, we designed eight primer sets and tested their abilities to amplify targets from representatives of each dicot and one monocot subclass. Five of the eight primer sets amplified targets from at least five of the seven taxa and thus exhibited broad taxonomic usefulness; the remaining primers were rather specific, however, and amplified targets from at most three taxa. In only one primer‐taxon combination was a complex multiple‐banded amplification produced. Overall, the protocol outlined proved quite useful at identifying broadly applicable primers targeted to low copy‐number nuclear genes. Wider application of this approach should be effective at greatly increasing the amount of genetic information available for a diversity of plant nuclear genomes.