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JXB Advance Access first published online on July 26, 2006
This version published online on July 31, 2006
Journal of Experimental Botany, doi:10.1093/jxb/erl065
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received March 20, 2006
Accepted May 23, 2006

RESEARCH PAPER

HaloTagTM: a new versatile reporter gene system in plant cells

Christina Lang 1, Jutta Schulze 1, Ralf-R. Mendel 1, and Robert Hänsch 1 *

1 Institut für Pflanzenbiologie, Technische Universität Braunschweig, Humboldtstraße 1, D-38106 Braunschweig, Germany

* To whom correspondence should be addressed.
Robert Hänsch, E-mail: r.haensch{at}tu-bs.de


  Abstract

HaloTagTM Interchangeable Labeling Technology (HaloTag) was originally developed for mammalian cell analysis. In this report, the use of HaloTag is demonstrated in plant cells for the first time. This system allows different fluorescent colours to be used to visualize the localization of the non-fluorescent HaloTag protein within living cells. A vector was constructed which expresses the HaloTag protein under the control of the 35S promoter of cauliflower mosaic virus. The functionality of the HaloTag construct was tested in transient assays by (i) transforming tobacco protoplasts and (ii) using biolistic transformation of intact leaf cells of tobacco and poplar plants. Two to fourteen days after transformation, the plant material was incubated with ligands specific for labelling the HaloTag protein, and fluorescence was visualized by confocal laser scanning microscopy. The results demonstrate that HaloTag technology is a flexible system which generates efficient fluorescence in different types of plant cells. The ligand-specific labelling of HaloTag protein was not hampered by the plant cell wall.

Keywords: cLSM; HaloTagTM; particle bombardment; protoplasts; reporter gene expression.

This version includes colour figures.


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