Author Liu Zijie Weng Yaguang, Li Suyan, Shi Qiong, Cai Yan, Liu Bin, Zhang Yan Yan Chen
Mechanism: Key Laboratory of Clinical Laboratory Diagnostics, Department of Medical Laboratory, Chongqing Medical University, Ministry of Education, Chongqing Key Laboratory, Chongqing 416
china biotechnology, vol.29, No.4, 29, pp.28-34
Keywords CENP-E Mps1 protein interaction fluorescence * * * vibrational energy transfer
Abstract Objective: To explore the CENP-E protein domain that interacts with Mps1 protein. Methods: human embryonic kidney 293(HEK293) cells were transfected with recombinant plasmids EGFP-CENPE2 (amino acids at positions 674-185
) and EGFP-CENPE3 (amino acids at positions 12-2134), and the receptor bleaching fluorescence * * * vibration energy transfer method was used (FRET's method
). Results: After transfection of HEK293 cells with plasmid with the weight < P >, it was observed by laser * * * focusing microscope that the fusion protein expressed by recombinant plasmid had * * * localization with Mps1. The FRET test results showed that the energy transfer rate between EGFP-CENPE3 and
Mps1 was [(12.63 .48)%, n = 3], and that between PEGFP-cenpe2 and Mps1 was
[(3.7 .21)%, n = 3], which was different from the control group. N=3] Comparing the energy transfer rates between pEGFP-CENPE3 and Mps1, there was a significant difference (P < .5). The results of immune * * * precipitation experiment showed that there was an interaction between EGFP-CENPE3 and Mps1 protein. Conclusion: FRET technique and immune * * * precipitation experiment prove that there is an interaction between
EGFP-CENPE3 and Mps1.