Yeast Two-Hybrid Screening to Test for Protein–Protein Interactions in the Auditory System

  • Dhasakumar S. Navaratnam
Part of the Methods in Molecular Biology™ book series (MIMB, volume 493)


We describe a protocol to screen for protein-protein interactions using the Gal-4 based yeast two-hybrid system. In this protocol, we describe serial transformation of bait into an already constructed cDNA library in yeast AH109 cells. We find this method to gives the most number of true interactions. Where a pre-made library in yeast cells is not available, the method outlined can be quickly adapted. AH109 cells can be first transformed with bait containing a vector followed by selection of yeast containing the bait. A second transformation of yeast cells is then accomplished with the cDNA library. The method is quick and can lead to the discovery of significant interactions.


Protein-protein interactions Yeast two-hybrid Gal4 based cochlea cDNA library pGBKT7 pGADT7 AH109 serial transformation 


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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Dhasakumar S. Navaratnam
    • 1
  1. 1.Department of NeurologyYale School of MedicineNew HavenUSA

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