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Journal
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Biochemistry
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RING-E3型无二体气体在Vitro和Planta中的功能特性
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal
Biochemistry
Functional Characterization of RING-Type E3 Ubiquitin Ligases In Vitro and In Planta
Please note that all translations are automatically generated.
Click here for the English version.
RING-E3型无二体气体在Vitro和Planta中的功能特性
DOI:
10.3791/60533-v
•
10:27 min
•
December 05, 2019
•
Joanna Kud
,
Wenjie Wang
,
Yulin Yuan
,
Allan Caplan
,
Joseph C. Kuhl
,
Louise-Marie Dandurand
,
Fangming Xiao
1
Department of Entomology, Plant Pathology and Nematology
,
University of Idaho
,
2
Department of Plant Sciences
,
University of Idaho
Chapters
00:04
Title
01:04
Site-directed Mutagenesis
02:29
Recombinant Protein Purification and In Vitro Ubiquitination Assay
03:40
Agrobacterium-mediated Transient Protein Expression in
Nicotiana benthamiana
Leaves and In Planta Ubiquitination Assay
06:36
Establishing the Link Between Enzymatic Activity and Function In Planta
07:27
Results: In Vitro and In Planta Ubiquitination Assay
09:20
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
本手稿的目的是为 RING 型 E3 泛性白化酶的综合生化和功能研究提供大纲。此多步骤管道,具有详细的协议,验证被测试蛋白质的酶活性,并演示如何将活动与功能联系起来。
Tags
RING-type E3 Ubiquitin Ligases
Site-directed Mutagenesis
In Vitro Enzymatic Activity
In Planta Functionality
Ubiquitination Assay
RING Domain
Cys And His Residues
PCR
Dpn1 Digestion
PMAL-c2 Vector
MBP Tag
SDS-PAGE
PVDF Membrane
Western Blotting
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