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褐藻外果藻基因组中因果基因座的定位:基于二代测序的定位和图位克隆方法

Localization of causal locus in the genome of the brown macroalga Ectocarpus: NGS-based mapping and positional cloning approaches.

作者信息

Billoud Bernard, Jouanno Émilie, Nehr Zofia, Carton Baptiste, Rolland Élodie, Chenivesse Sabine, Charrier Bénédicte

机构信息

Sorbonne Université, UPMC University Paris 06, CNRS, UMR 8227, Integrative Biology of Marine Models, Station Biologique de Roscoff CS 90074, F-29688, Roscoff cedex, France.

出版信息

Front Plant Sci. 2015 Feb 19;6:68. doi: 10.3389/fpls.2015.00068. eCollection 2015.

Abstract

Mutagenesis is the only process by which unpredicted biological gene function can be identified. Despite that several macroalgal developmental mutants have been generated, their causal mutation was never identified, because experimental conditions were not gathered at that time. Today, progresses in macroalgal genomics and judicious choices of suitable genetic models make mutated gene identification possible. This article presents a comparative study of two methods aiming at identifying a genetic locus in the brown alga Ectocarpus siliculosus: positional cloning and Next-Generation Sequencing (NGS)-based mapping. Once necessary preliminary experimental tools were gathered, we tested both analyses on an Ectocarpus morphogenetic mutant. We show how a narrower localization results from the combination of the two methods. Advantages and drawbacks of these two approaches as well as potential transfer to other macroalgae are discussed.

摘要

诱变是唯一能够识别未预测到的生物基因功能的过程。尽管已经产生了几种大型海藻发育突变体,但由于当时没有收集实验条件,它们的因果突变从未被确定。如今,大型海藻基因组学的进展以及对合适遗传模型的明智选择使得识别突变基因成为可能。本文对旨在鉴定褐藻硅藻(Ectocarpus siliculosus)中一个基因座的两种方法进行了比较研究:定位克隆和基于下一代测序(NGS)的作图。一旦收集到必要的初步实验工具,我们就在一种硅藻形态发生突变体上测试了这两种分析方法。我们展示了两种方法的结合如何产生更窄的定位结果。讨论了这两种方法的优缺点以及向其他大型海藻转移的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f05a/4333798/9b93d5b2d24e/fpls-06-00068-g0001.jpg

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