Preparation and electroporation of oat protoplasts from cell suspension culture.
Preparation and electroporation of oat protoplasts from cell suspension culture.
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细胞悬浮培养物燕麦原生质体的制备和电穿孔。
DOI：
10.1002/9780471729259.mc16d03s05
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发表时间：
2007
影响因子：
--
通讯作者：
Miller,WAllen
中科院分区：
文献类型：
--
作者：
Rakotondrafara,AurelieM;Jackson,JacquelynR;Kneller,ElizabethPettit;Miller,WAllen
原生质体为将核酸导入植物细胞提供了一个方便的系统。原生质体允许快速测定基因表达和病毒复制，对于植物生物学具有与用于动物系统研究的培养动物细胞相似的优势。传统上，原生质体的制备和处理既是科学又是艺术，需要使用者的特殊操作。本单元的目的是清楚详细地介绍原生质体制备中涉及的所有方法和细微差别，使用一个强大，可靠的系统，不需要超出非专业分子生物学家预期的技能。由于双子叶植物和单子叶植物在许多生物学特性上存在差异，而且不同的方法可能对不同的植物更有效，因此本章将分别介绍双子叶植物（拟南芥、烟草;参见单元16D.1和16D.4）和单子叶植物（燕麦）的原生质体制备。本单元描述了燕麦悬浮培养物原生质体的制备和电穿孔转染方法。
Protoplasts provide a convenient system for introduction of nucleic acids into plant cells. Protoplasts allow rapid assays of gene expression and virus replication, with advantages for plant biology similar to those of cultured animal cells for the study of animal systems. Traditionally, preparation and handling of protoplasts has been as much art as science, requiring a special touch by the user. The purpose of this unit is to lay out in clear detail all the methods and nuances involved in protoplast preparation using a robust, reliable system that does not require skills beyond those expected of an unspecialized molecular biologist. Because dicots and monocots differ in many biological properties, and because different procedures may work better for different plants, separate units in this chapter are devoted to protoplast preparation from dicots (Arabidopsis, tobacco; refer toUNITS 16D.1 & 16D.4) and from a monocot (oat). This unit describes methods for preparation and transfection by electroporation of protoplasts derived from an oat suspension culture.