Bulked-Segregant Analysis Coupled to Whole Genome Sequencing (BSA-Seq) for Rapid Gene Cloning in Maize.
Bulked-Segregant Analysis Coupled to Whole Genome Sequencing (BSA-Seq) for Rapid Gene Cloning in Maize.
复制标题
批量隔离分析与整个基因组测序（BSA-SEQ）耦合，以在玉米中快速基因克隆。
DOI：
10.1534/g3.118.200499
复制
发表时间：
2018-11-06
期刊：
影响因子：
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通讯作者：
Bartlett M
中科院分区：
文献类型：
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作者：
Klein H;Xiao Y;Conklin PA;Govindarajulu R;Kelly JA;Scanlon MJ;Whipple CJ;Bartlett M
正向遗传学仍然是揭示支撑有机体形式和功能的基因以及揭示这些基因如何在基因网络中联系在一起的强大方法。在玉米中，正向遗传学取得了巨大成功，但玉米基因组的大小和复杂性使得用传统方法识别突变基因往往是一个艰巨的过程。下一代测序革命使得许多生物体的基因克隆过程显着加速，即使基因组庞大且复杂。在这里，我们描述了用于克隆玉米突变基因的批量分离分析测序（BSA-Seq）方案。我们的简单策略可用于从汇集的 F2 个体的全基因组测序中快速识别作图区间和候选单核苷酸多态性 (SNP)。我们采用这种策略来识别窄奇矮星作为大蜀黍分支1的增强子，并识别有缺陷的kernel1的新等位基因。我们的方法提供了一种快速、简单的方法来克隆玉米基因。
Forward genetics remains a powerful method for revealing the genes underpinning organismal form and function, and for revealing how these genes are tied together in gene networks. In maize, forward genetics has been tremendously successful, but the size and complexity of the maize genome made identifying mutant genes an often arduous process with traditional methods. The next generation sequencing revolution has allowed for the gene cloning process to be significantly accelerated in many organisms, even when genomes are large and complex. Here, we describe a bulked-segregant analysis sequencing (BSA-Seq) protocol for cloning mutant genes in maize. Our simple strategy can be used to quickly identify a mapping interval and candidate single nucleotide polymorphisms (SNPs) from whole genome sequencing of pooled F2 individuals. We employed this strategy to identify narrow odd dwarf as an enhancer of teosinte branched1, and to identify a new allele of defective kernel1. Our method provides a quick, simple way to clone genes in maize.