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. Author manuscript; available in PMC: 2015 Jul 15.
Published in final edited form as: Methods Mol Biol. 2013;977:323–338. doi: 10.1007/978-1-62703-284-1_26

Table 1.

Experimental Design for Testing Protein:Protein Interactions

Sample ACT Vector BIND vector pGL4.31
1 pFN10A (ACT)-Y pFN11A (BIND)-X +
2 pACT Vector pFN11A (BIND)-X +
3 pFN10A (ACT)-Y pBIND Vector +
4 pACT Vector pBIND Vector +
5 pACT-MyoD Control pBIND-Id Control +

Sample 1 is used to test for protein interactions between the X and Y protein-coding regions. Sample 2 is used to test for the background level of reporter activity when one of the protein-coding regions is fused to the GAL4 DNA-binding domain. This also tests for transcriptional activation domain activity of protein X when fused to the GAL4 DNA-binding domain. Sample 3 is used to test for the background levels of reporter activity when one of the protein-coding regions is fused to the activation domain. Sample 4 generally provides the lowest level of background activity of luciferase activity from the reporter plasmid. Sample 5 provides a positive control with interacting proteins fused to the activation and GAL4 DNA-binding domains