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Archive > Volume 25 Issue 3 > 2018,25(3):258-262. DOI:10.3872/j.issn.1007-385X.2018.03.008 Prev Next
Basic Research
Screening of S-phase kinase-assuciated protein 2 in the cervical carcinoma Hela cell
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Abstract:
Objective： ：The co-immunoprecipitation and mass spectrometric analysis was carried out to obtain the S-phase kinase-asso- ciated protein 2 (SKP2)-binding proteins in HeLa cells, and the biological functions of these binding proteins were forecast. Methods: The co-immunoprecipitation system was established by co-immunoprecipitation and Western blotting assay; the specific protein gel of SKP2-binding proteins was obtained by SDS-PAGE and silver staining assay; the potential SKP2-binding proteins was identified by mass spectrometric analysis; and the GO (Gene ontology) analysis and KEGG analysis was carried out by bioinformatics technique. Results: The expression level of SKP2 protein in HeLa cells was high enough for co-immunoprecipitation assay; the co-immunoprecipi- tation system was established successfully, and SKP2-binding proteins was obtained; a total of 563 proteins were identified by mass spectrometric analysis, and 270 proteins with high credibility were obtained after screening. The GO analysis and KEGG analysis was carried out for the 270 proteins to forecast their functions and pathways. Conclusion: The SKP2-binding proteins were screened suc- cessfully, and it was the foundation for the subsequent screening of target-binding proteins and the search for targeting drugs.
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Project Supported:
Project supported by the Natural Science Foundation of Zhejiang Province(No.LY18H160067，LQ12H16013)，and the Zhejiang Provincial Medical and Healthy Foundation for Science and Technology Program(No.2015KYB090，2011KYB005)
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