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

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 spec...

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Autores principales: Román, Ángela, Golz, John F., Webb, Alex A. R., Graham, Ian A., Haydon, Michael J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7217008/
https://www.ncbi.nlm.nih.gov/pubmed/31692146
http://dx.doi.org/10.1111/tpj.14603
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author Román, Ángela
Golz, John F.
Webb, Alex A. R.
Graham, Ian A.
Haydon, Michael J.
author_facet Román, Ángela
Golz, John F.
Webb, Alex A. R.
Graham, Ian A.
Haydon, Michael J.
author_sort Román, Ángela
collection PubMed
description 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.
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spelling pubmed-72170082020-05-13 Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis Román, Ángela Golz, John F. Webb, Alex A. R. Graham, Ian A. Haydon, Michael J. Plant J Technical Advance 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. John Wiley and Sons Inc. 2019-12-03 2020-04 /pmc/articles/PMC7217008/ /pubmed/31692146 http://dx.doi.org/10.1111/tpj.14603 Text en © 2019 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Technical Advance
Román, Ángela
Golz, John F.
Webb, Alex A. R.
Graham, Ian A.
Haydon, Michael J.
Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis
title Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis
title_full Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis
title_fullStr Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis
title_full_unstemmed Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis
title_short Combining GAL4 GFP enhancer trap with split luciferase to measure spatiotemporal promoter activity in Arabidopsis
title_sort combining gal4 gfp enhancer trap with split luciferase to measure spatiotemporal promoter activity in arabidopsis
topic Technical Advance
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7217008/
https://www.ncbi.nlm.nih.gov/pubmed/31692146
http://dx.doi.org/10.1111/tpj.14603
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