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. 2010 Aug 26;38(20):e189. doi: 10.1093/nar/gkq757

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© The Author(s) 2010. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), 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. — Integration vectors to construct one-hybrid strains with ADE2 and HIS3 reporter genes. (A) Integration vector pOHAK for ADE2 reporter gene. We cloned a fragment spanning AFI1-GAL2_UAS-GAL2_TATA-ADE2 from PJ69-2A and replaced the GAL2_UAS with a fragment containing KanMX and an HpaI site to obtain pOHAK. (B) Integration vector pOHHZ for HIS3 reporter gene. We cloned a fragment spanning LYS2-GAL1_UAS-GAL1_TATA-HIS3 from PJ69-2A and replaced the GAL1_UAS with a fragment containing Zeo^R and a PmeI site to obtain pOHHZ. (C) Schematic representation of reporter gene construction in the case of ADE2 as an example. A methylatable bait sequence is cloned at the HpaI site of pOHAK, and the plasmid linearized by SalI-KpnI digestion is used for transformation of PJ69-2A. Homologous recombination occurs within ORFs for AFI1 and ADE2.