Tetracycline operator/repressor system to visualize fluorescence-tagged T-DNAs in interphase nuclei ofArabidopsis View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2003-03

AUTHORS

Antonius J. M. Matzke, Johannes van der Winden, Marjori Matzke

ABSTRACT

The bacterial tetracycline operator/repressor (tetO/TetR) system and enhanced yellow fluorescent protein (EYFP) have been adapted for use inArabidopsis to visualize tagged T-DNAs in interphase nuclei of living cells. The 2-component system was assembled on a single T-DNA construct that contained a gene encoding a nuclear-targeted TetR-EYFP fusion protein under the control of the 35S promoter of the cauliflower mosaic virus together with multiple tandemly arranged copies of thetetO. In a number of independentArabidopsis lines transformed with this construct, bright fluorescent dots corresponding to tagged T-DNAs were observed in interphase nuclei of various cell types using standard fluorescence microscopy. In selfed progeny of a single locus line, hemizygous and homozygous plants were distinguished by having 1 or 2 fluorescent dots, respectively. Low background fluorescence of EYFP in many plant tissues facilitates the visualization of tagged T-DNAs. We compared features of thetetO/TetR-EYFP system to a second system we developed on the basis of the bacteriallac operator/repressor and enhanced green fluorescent protein. More... »

PAGES

9-19

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf02773392

DOI

http://dx.doi.org/10.1007/bf02773392

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1051010080


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