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. 2016 Sep 22;6:32445. doi: 10.1038/srep32445

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Copyright © 2016, The Author(s)

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Expression verification of the bait fusion vector (pDHB1 + P10) (A) pDHB1 + P10 bait fusion vector confirmation by restriction enzyme digestion. Lane M: Lambda DNA/HindIII marker, lane 1: digestion of pDHB1 bait vector by enzyme, lane 2: digestion of pDHB1 + P0 bait fusion vector by enzyme, lane 3: P10, M’: DL 2,000 marker. (B) Detection of pDHB1 + P10 bait fusion protein in western blot. Lane 1: pDHB1 + P10, lane 2: pPR3-N empty insert (negative control). (C) Functional analysis of pDHB1 + P10 bait protein in yeast. Bait fusion vector (pDHB1 + P10) was used to cotransform yeast with the control plasmid pOst1-NubI or pPR3-N empty insert and grown on selective SD medium. Coexpression of P10 with Ost1-NubI resulted in growth of yeast transformants on selective media as a sign of reporter gene activation. Coexpression of P10 with pPR3-N empty insert did not yield any colonies on selective medium. pDHB1-LargeT and pDSL-p53 were used as positive controls, pDHB1-LargeT and pPR3-N-empty insert were used as negative controls.