Plant Y1H/Y2H Point-to-Point Validation Service
Candidate Interaction Validation in Yeast
CD BioSciences provides Plant Y1H/Y2H point-to-point validation services for testing defined DNA-protein or protein-protein candidate interactions. The service is suitable for follow-up validation after library screening or for direct testing of known candidate pairs.
Point-to-point validation can include bait and prey vector construction, auto-activation and toxicity assessment, yeast co-transformation, selective growth, dilution spotting, imaging, and result interpretation.
When to Use Point-to-Point Validation
Re-test selected Y1H or Y2H hits in a defined bait-prey combination.
Evaluate candidate transcription factors, promoter fragments, or protein pairs selected from prior data.
Compare full-length, domain-truncated, or motif-mutated constructs.
Assess bait background, prey behavior, and selection conditions before larger screening.
Validation Designs
Y1H point-to-point validation tests whether a candidate prey protein can bind a defined DNA bait in a yeast reporter system. Y2H point-to-point validation tests whether a defined bait protein and prey protein interact in a yeast reporter system. Both designs require background controls to avoid overinterpreting self-activation or non-specific growth.
Figure 1. Point-to-point validation design for Y1H and Y2H candidate interactions.
Controls
| Control | Purpose |
|---|---|
| Bait-only control | Detects self-activation or background growth caused by the bait construct. |
| Prey-only or empty vector control | Helps evaluate prey-related reporter activation or non-specific growth. |
| Positive control | Confirms that the yeast system and selective plates support expected reporter activation. |
| Negative control | Provides a baseline for non-interacting combinations. |
| Dilution spotting | Compares yeast growth under selected stringency conditions. |
Workflow
Figure 2. Representative dilution spotting strategy for yeast hybrid validation.
- Review candidate pair or bait-prey sequence information
- Construct bait and prey vectors
- Perform bait and prey background checks when required
- Co-transform yeast cells with defined construct combinations
- Apply selective media and dilution spotting
- Image plates and interpret growth patterns
- Prepare report and original image files
Interpretation Notes
Growth on selective plates supports interaction under yeast assay conditions, but it does not by itself establish in planta interaction. For high-confidence conclusions, yeast validation is often combined with EMSA, dual-luciferase, Co-IP, GST pull-down, BiFC, LCA, or related assays depending on the biological question.
Sample and Information Requirements
- Candidate DNA bait or protein bait sequence
- Candidate prey gene or protein sequence
- Existing plasmid information, if available
- Expected Y1H or Y2H validation design
- Number of candidate pairs
- Preferred follow-up validation method, if any
Deliverables
- Vector construction information
- Auto-activation or toxicity test record, if performed
- Co-transformation and spotting images
- Positive/negative control result summary
- Result interpretation and standard report
Related Services
Please contact us with your candidate bait and prey sequences, validation goal, and number of pairs for project evaluation.
For research use only, not for clinical use.