Journal
PLANT SCIENCE
Volume 274, Issue -, Pages 2-7Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.plantsci.2018.03.032
Keywords
Biolistic transient expression; Seedling lethal mutant; Juvenile rice; Protein subcellular localization and dynamics
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Funding
- Research Grants Council of Hong Kong [CUHK14130716, CUHK14102417, C4011-14R, C4012-16E, C4002-17G, AoE/M-05/12]
- National Natural Science Foundation of China [31270226, 31470294]
- Research Committee of CUHK
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Knowledge on the subcellular localization of target proteins in a plant mutant background is important for revealing the function of the genes investigated. However, in Arabidopsis and rice, mutant lethality is one major barrier to such studies. Here we describe an optimized bombardment-mediated transient expression approach for studying subcellular protein localization in Arabidopsis seedling of lethal mutants. The whole experiment comprises four stages: cultivation and preparation of plants, coating gold particles with plasmid DNA, delivery of DNA into plants via bombardment, plant incubation and gene expression analysis which include localization and dynamics, co-localization comparison with reporter proteins and functional analysis. The entire process takes about 3-10 days from plant cultivation to protein detection. It has a high efficiency and the results are reproducible. Additionally, this protocol is applicable for the transient expression of chimeric fluorescent fusion proteins in juvenile rice seedlings and leaf sheaths, saving time dramatically in comparison of generating transgenic rice plant.
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