一种分离大豆原生质体并应用于瞬时基因表达分析的简单方法。
A Simple Method for Isolation of Soybean Protoplasts and Application to Transient Gene Expression Analyses.
作者信息
Wu Faqiang, Hanzawa Yoshie
机构信息
Department of Biology, California State University, Northridge.
Department of Biology, California State University, Northridge;
出版信息
J Vis Exp. 2018 Jan 25(131):57258. doi: 10.3791/57258.
Abstract
Soybean (Glycine max (L.) Merr.) is an important crop species and has become a legume model for the studies of genetic and biochemical pathways. Therefore, it is important to establish an efficient transient gene expression system in soybean. Here, we report a simple protocol for the preparation of soybean protoplasts and its application for transient functional analyses. We found that young unifoliate leaves from soybean seedlings resulted in large quantities of high quality protoplasts. By optimizing a PEG-calcium-mediated transformation method, we achieved high transformation efficiency using soybean unifoliate protoplasts. This system provides an efficient and versatile model for examination of complex regulatory and signaling mechanisms in live soybean cells and may help to better understand diverse cellular, developmental and physiological processes of legumes.
摘要
大豆（Glycine max (L.) Merr.）是一种重要的作物物种，已成为用于研究遗传和生化途径的豆科植物模型。因此，在大豆中建立高效的瞬时基因表达系统很重要。在此，我们报告了一种制备大豆原生质体的简单方案及其在瞬时功能分析中的应用。我们发现，大豆幼苗的幼嫩单叶能产生大量高质量的原生质体。通过优化聚乙二醇-钙介导的转化方法，我们利用大豆单叶原生质体实现了高转化效率。该系统为研究活大豆细胞中复杂的调控和信号传导机制提供了一个高效且通用的模型，可能有助于更好地理解豆科植物的各种细胞、发育和生理过程。
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