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231223s2011 xx |||||o 00| ||eng c |
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|a 10.1111/j.1469-8137.2010.03587.x
|2 doi
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|a pubmed25n0682.xml
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|a DE-627
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|e rakwb
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|a eng
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| 100 |
1 |
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|a Buschmann, H
|e verfasserin
|4 aut
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|a Cytoskeletal dynamics in interphase, mitosis and cytokinesis analysed through Agrobacterium-mediated transient transformation of tobacco BY-2 cells
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|c 2011
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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| 500 |
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|a Date Completed 31.12.2014
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|a Date Revised 13.12.2023
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a © 2010 The Authors. New Phytologist © 2010 New Phytologist Trust.
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|a Transient transformation with Agrobacterium is a widespread tool allowing rapid expression analyses in plants. However, the available methods generate expression in interphase and do not allow the routine analysis of dividing cells. Here, we present a transient transformation method (termed 'TAMBY2') to enable cell biological studies in interphase and cell division. Agrobacterium-mediated transient gene expression in tobacco BY-2 was analysed by Western blotting and quantitative fluorescence microscopy. Time-lapse microscopy of cytoskeletal markers was employed to monitor cell division. Double-labelling in interphase and mitosis enabled localization studies. We found that the transient transformation efficiency was highest when BY-2/Agrobacterium co-cultivation was performed on solid medium. Transformants produced in this way divided at high frequency. We demonstrated the utility of the method by defining the behaviour of a previously uncharacterized microtubule motor, KinG, throughout the cell cycle. Our analyses demonstrated that TAMBY2 provides a flexible tool for the transient transformation of BY-2 with Agrobacterium. Fluorescence double-labelling showed that KinG localizes to microtubules and to F-actin. In interphase, KinG accumulates on microtubule lagging ends, suggesting a minus-end-directed function in vivo. Time-lapse studies of cell division showed that GFP-KinG strongly labels preprophase band and phragmoplast, but not the metaphase spindle
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a Agrobacterium
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|a cell division
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|a cytoskeleton
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|a kinesin
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|a microtubule dynamics
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| 650 |
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4 |
|a tobacco BY-2
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|a transient gene expression
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|a Kinesins
|2 NLM
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| 650 |
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|a EC 3.6.4.4
|2 NLM
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| 700 |
1 |
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|a Green, P
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Sambade, A
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Doonan, J H
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Lloyd, C W
|e verfasserin
|4 aut
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| 773 |
0 |
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|i Enthalten in
|t The New phytologist
|d 1990
|g 190(2011), 1 vom: 01. Apr., Seite 258-267
|w (DE-627)NLM09818248X
|x 1469-8137
|7 nnns
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| 773 |
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|g volume:190
|g year:2011
|g number:1
|g day:01
|g month:04
|g pages:258-267
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|u http://dx.doi.org/10.1111/j.1469-8137.2010.03587.x
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