%A LIU Yong-mei, TAN Ming, LEI Ming %T Electron microscopic study of the human THOC1-THOC2-THOC3 subcomplex %0 Journal Article %D 2020 %J Journal of Shanghai Jiao Tong University (Medical Science) %R 10.3969/j.issn.1674-8115.2020.01.001 %P 1- %V 40 %N 1 %U {https://xuebao.shsmu.edu.cn/CN/abstract/article_12476.shtml} %8 2020-01-28 %X 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.