Use of Maize (Zea mays L.) Mutator Transposon-Induced Mutants of the BonnMu Resource for Forward and Reverse Genetics Studies.

Yan Naing Win, Martin Pöschel, Tyll Stöcker, Xuelian Du, Alina Klaus, Ben Wilhelm Braun, Linnéa Lukas, Alexa Brox, Heiko Schoof, Frank Hochholdinger, Caroline Marcon
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Abstract

The BonnMu resource represents a tagged collection of maize (Zea mays L.) Mutator (Mu) transposon-induced mutants, designed for functional genomics studies. Here, we describe the use of the BonnMu collection for identifying and characterizing mutations. Specifically, we describe workflows for use in both reverse and forward genetics strategies in maize. For reverse genetics, users first acquire a BonnMu F2 stock of interest based on data accessible at the Maize Genetics and Genomics Database (MaizeGDB). We provide details here for their subsequent propagation and for the confirmation of Mu insertions by genotyping via PCR, with the ultimate goal of establishing genotype-phenotype relationships of interest. For forward genetics studies, we describe a workflow that involves a combined approach of Mutant-Seq (Mu-Seq) and bulked segregant RNA-seq (BSR-Seq), to identify the causal gene underlying a mutant phenotype of interest.

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利用波恩姆资源的玉米(Zea mays L.)突变体转座子诱导突变体进行正向和反向遗传学研究。
BonnMu 资源是玉米(Zea mays L.)Mutator(Mu)转座子诱导突变体的标记集合,专为功能基因组学研究而设计。在此,我们将介绍如何使用 BonnMu 收集来鉴定和表征突变。具体来说,我们介绍了用于玉米反向和正向遗传学策略的工作流程。在反向遗传学中,用户首先要根据玉米遗传学和基因组学数据库(MaizeGDB)中的数据获取感兴趣的波恩穆 F2 种群。在此,我们将详细介绍如何进行后续繁殖,以及如何通过 PCR 基因分型确认 Mu 插入,最终目标是建立感兴趣的基因型-表型关系。在前向遗传学研究方面,我们介绍了一种工作流程,其中包括突变体-序列(Mu-Seq)和大量分离 RNA-序列(BSR-Seq)的组合方法,以确定突变体表型背后的致病基因。
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Cold Spring Harbor protocols
Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
3.00
自引率
0.00%
发文量
163
期刊介绍: Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.
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