Novel interactions of the von Hippel-Lindau (pVHL) tumor suppressor with the CDKN1 family of cell cycle inhibitors
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| Peptide Synthesis |
The pcDNA3.1-derived plasmids carrying synthetic full-length cDNA sequences coding for the human pVHL30, p21, p27 and p57 proteins were purchased from GenScript (GenEZ plasmids OHu23297, OHu27895, OHu26670 and OHu27234 respectively).All recombinant plasmids used in the yeast two-hybrid assays (Supplementary Table S1) are able to express the proteins of interest in fusion with either the DNA binding domain (DBD) or the activation domain (AD) of the Gal4 transcription factor. The cMyc and HA epitopes present in the fusion proteins with the Gal4 DBD and AD, respectively, allow to readily verify the expression of the chimeric proteins in yeast cells. GenScript plasmids have been directly used to overexpress the different CDKN1 proteins as N-terminal FLAG-tagged fusion polypeptides in mammalian cells. |
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