TY - JOUR
T1 - Re-assessment of biolistic transient expression
T2 - An efficient and robust method for protein localization studies in seedling-lethal mutant and juvenile plants
AU - Wang, Hao
AU - Zhao, Qiong
AU - Fu, Jiaxin
AU - Wang, Xiangfeng
AU - Jiang, Liwen
N1 - Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/9
Y1 - 2018/9
N2 - 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.
AB - 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.
KW - Biolistic transient expression
KW - Juvenile rice
KW - Protein subcellular localization and dynamics
KW - Seedling lethal mutant
UR - https://www.scopus.com/pages/publications/85045541708
U2 - 10.1016/j.plantsci.2018.03.032
DO - 10.1016/j.plantsci.2018.03.032
M3 - 文章
C2 - 30080604
AN - SCOPUS:85045541708
SN - 0168-9452
VL - 274
SP - 2
EP - 7
JO - Plant Science
JF - Plant Science
ER -