Journal of Shanghai Jiao Tong University (Medical Science) >

2020 , Vol. 40 >Issue 1: 1

DOI: https://doi.org/10.3969/j.issn.1674-8115.2020.01.001

Original article (Basic research)

Electron microscopic study of the human THOC1-THOC2-THOC3 subcomplex

  • LIU Yong-mei ,
  • TAN Ming ,
  • LEI Ming
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  • Shanghai Institute of Precision Medicine, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China

Online published: 2020-03-05

Supported by

National Natural Science Foundation of China (31525007).

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Abstract

Objective · To study the structure of the human THOC1-THOC2-THOC3 subcomplexnegative-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 followedSuperose 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 obtainedusing a tandem affinity purification scheme with Ni-NTA and streptavidin-based chromatography. Preliminary study on the structure of human THOC1-THOC2-THOC3 subcomplex was achievedelectron microscopy using negative staining with uranium formate. Conclusion · A low resolution model of human THOC1-THOC2-THOC3 subcomplex was achievedsingle particle reconstruction.

Key words： mRNA nuclear transport; TREX complex; THO complex; negative-staining electron microscopy

Cite this article

LIU Yong-mei , TAN Ming , LEI Ming . Electron microscopic study of the human THOC1-THOC2-THOC3 subcomplex[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2020 , 40(1) : 1 . DOI: 10.3969/j.issn.1674-8115.2020.01.001

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