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Methods Mol Biol. 2016;1370:17-25. doi: 10.1007/978-1-4939-3142-2_2.

Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells.

Author information

1
Botany Department, School of Biology and Chemistry, Osnabrück University, Postfach 44 69, Osnabrück, 49069, Germany. Henrik.buschmann@biologie.uni-osnabrueck.de.

Abstract

The continuing analysis of plant cell division will require additional protein localization studies. This is greatly aided by GFP-technology, but plant transformation and the maintenance of transgenic lines can present a significant technical bottleneck. In this chapter I describe a method for the Agrobacterium-mediated genetic transformation of tobacco BY-2 cells. The method allows for the microscopic analysis of fluorescence-tagged proteins in dividing cells in within 2 days after starting a coculture. This transient transformation procedure requires only standard laboratory equipment. It is hoped that this rapid method would aid researchers conducting live-cell localization studies in plant mitosis and cytokinesis.

KEYWORDS:

Agrobacterium; Cell division; Cytokinesis; F-actin; Green fluorescent protein; Tobacco BY-2; Transient transformation

PMID:
26659951
DOI:
10.1007/978-1-4939-3142-2_2
[Indexed for MEDLINE]

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