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Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis.

Accepted version
Peer-reviewed

Type

Article

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Abstract

In multicellular organisms different types of tissues have distinct gene expression profiles associated with specific function or structure of the cell. Quantification of gene expression in whole organs or whole organisms can give misleading information about levels or dynamics of expression in specific cell types. Tissue- or cell-specific analysis of gene expression has potential to enhance our understanding of gene regulation and interactions of cell signalling networks. The Arabidopsis circadian oscillator is a gene network which orchestrates rhythmic expression across the day/night cycle. There is heterogeneity between cell and tissue types of the composition and behaviour of the oscillator. In order to better understand the spatial and temporal patterns of gene expression, flexible tools are required. By combining a Gateway®-compatible split luciferase construct with a GAL4 GFP enhancer trap system, we describe a tissue-specific split luciferase assay for non-invasive detection of spatiotemporal gene expression in Arabidopsis. We demonstrate the utility of this enhancer trap-compatible split luciferase assay (ETSLA) system to investigate tissue-specific dynamics of circadian gene expression. We confirm spatial heterogeneity of circadian gene expression in Arabidopsis leaves and describe the resources available to investigate any gene of interest.

Description

Keywords

Arabidopsis, circadian clock, enhancer trap, gene expression, luciferase, technical advance, tissue-specificity, Arabidopsis, Circadian Clocks, Gene Expression Regulation, Plant, Genes, Plant, Genetic Markers, Genetic Techniques, Luciferases, Plants, Genetically Modified, Polymerase Chain Reaction, Promoter Regions, Genetic

Journal Title

Plant J

Conference Name

Journal ISSN

0960-7412
1365-313X

Volume Title

102

Publisher

Wiley

Rights

All rights reserved
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/H006826/1)