Yeast Two-Hybrid Protein Interaction Screening Service
Yeast Two-Hybrid (Y2H) screening is a well-established method to detect protein protein interactions (see here for a powerpoint explanation of the basic method).
The DNA binding domain (DBD) of the transcription factor is expressed as a hybrid protein fused to the protein of interest (marked "B" as 'bait', see top left in the picture), and the activation domain (AD) is fused to another protein ("P" as 'prey', see top right in the picture).
If an interaction between bait and prey takes place the activation domain will come into the proper position to activate a reporter gene. In an Y2H screen a bait protein is tested against a complex collection of proteins or protein fragments originating from cDNA or ORF libraries (fusions with the AD). Interacting proteins (preys) are identified by sequencing the corresponding plasmid DNA. We offer more than 30 different cDNA screening libraries from several species and tissues and two human OFR libraries (N- and C-terminal constructs with >10,000 ORFs each).
For frequently asked questions, please see our FAQ-list.
What we need from you
We need your bait DNA cloned into a plasmid suitable for our system, such as pGBT9 or pGBKT7. pGBKCg or a Gateway-compatible variant. Small aliquots are available from us upon request. We require 10 µg DNA of your bait construct at a concentration > 100ng/µl.
What we do
The yeast two-hybrid screening service offers protein interaction screens with an automated high-throughput Y2H screening procedure. We
- Optimise screening conditions for your bait
- Screen libraries of your choice
- Retrieve the library inserts of positive clones by PCR
- Sequence the inserts from the 5' end
- Analyse the sequence by blast analysis
- Compare the results to previous screens
- Prepare a report for you
What we return
You will receive a report from us highlighting interacting proteins that qualify as high-confidence interactors based on comparisons with previous screens. You can also get the PCR products 'harbouring' the library inserts. We generally do not keep the positive yeast cells.
For more details regarding the two-hybrid service, cloning vectors, experimental planning support, or prices for external customers, please contact Sara Burmester. Information on pricing for internal customers can be found at the GPCF pricelist.
» Y2H-screening-service: Frequently Asked Questions
If you are interested in a pair-wise analysis of protein-protein interactions in a mammalian cell system, please have a look at our
