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. 2007 Nov 5;35(21):e142. doi: 10.1093/nar/gkm932

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© 2007 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Yeast two-hybrid system for the detection of pVHL–substrate interactions. (a) Essential elements of the system. The BD-pVHL bait and AD-HIF1α prey fusions are expressed from the pBridge and pACT2 vectors under the control of the constitutive P_ADH1 promoter. The pBridge vector in addition allows for the expression of PHD1, 2 or 3 under the control of the P_MET25 promoter, which is induced in the absence of methionine. The yeast reporter strain CG1945BC is a derivative of CG1945 expressing ELB and ELC under the control of the constitutive P_PGK1 promoter from chromosome XV next to the ADE2 locus (19). (b) Schematic representation of reporter gene activation by the PHD-, ELB-, ELC-dependent interaction between BD-pVHL and AD-HIF1α. Pro, HO-Pro; unmodified and hydroxylated form, respectively, of PHD target prolyl residues.