Electron microscopic study of the human THOC1-THOC2-THOC3 subcomplex / 上海交通大学学报(医学版)
Journal of Shanghai Jiaotong University(Medical Science)
;
(12): 1-7, 2020.
Article
in Chinese
| WPRIM
| ID: wpr-843255
ABSTRACT
Objective:
To study the structure of the human THOC1-THOC2-THOC3 subcomplex by negative-staining electron microscopy.Methods:
The recombinant protein components were co-expressed in cultured insect cells, and the protein complex was isolated via sequential Ni-NTA and Strep-Tactin affinity purification followed by Superose gel filtration chromatography. The purified protein sample was then subjected to negative-staining electron microscopy and single particle image analysis.Results:
The recombinant human THOC1-THOC2-THOC3 complex with high purity and good homogeneity was obtained by using a tandem affinity purification scheme with Ni-NTA and streptavidin-based chromatography. Preliminary study on the structure of human THOC1-THOC2-THOC3 subcomplex was achieved by electron microscopy using negative staining with uranium formate.Conclusion:
A low resolution model of human THOC1-THOC2-THOC3 subcomplex was achieved by single particle reconstruction.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Journal of Shanghai Jiaotong University(Medical Science)
Year:
2020
Type:
Article
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