Plant Nuclear Yeast Two-Hybrid Screening Service
Conventional Y2H Screening for Soluble Plant Proteins
CD BioSciences provides Plant Nuclear Yeast Two-Hybrid screening services for identifying candidate interactors of soluble or nuclear-compatible plant bait proteins.
This service focuses on conventional GAL4-based or related nuclear reporter Y2H systems, where bait and prey fusion proteins interact in yeast and activate reporter gene expression.
Principle
In nuclear Y2H, the bait protein is fused to a DNA-binding domain and the prey protein library is fused to a transcriptional activation domain. Interaction between bait and prey reconstitutes transcriptional activation of reporter genes, allowing growth or color-based screening under selective conditions.
Figure 1. Conventional nuclear Y2H reporter principle.
Suitable Bait Proteins
Proteins without strong transmembrane regions or membrane topology constraints.
Candidate regulators, transcription factors, enzymes, adaptors, and signaling proteins.
Selected domains or truncated regions can be tested to reduce self-activation or improve expression.
Specific bait-prey combinations can be tested by point-to-point nuclear Y2H validation.
Screening Workflow
Figure 2. Nuclear Y2H screening and positive clone confirmation workflow.
- Bait sequence review and construct planning
- Bait vector construction
- Self-activation and toxicity testing
- Prey library construction or selection
- Library screening under selective conditions
- Positive clone PCR and sequencing
- Hit annotation and report preparation
Self-activation and Bait Design
Some plant proteins, especially transcription factors or activation-domain-containing proteins, may activate reporter genes without a prey partner. If self-activation occurs, bait truncation, domain redesign, reporter stringency adjustment, or an alternative method may be considered.
| Issue | Possible Response |
|---|---|
| Strong self-activation | Evaluate truncation, domain deletion, or alternative bait design. |
| Bait toxicity | Adjust construct design or expression context where feasible. |
| Poor transformant recovery | Review plasmid quality, construct size, and yeast transformation conditions. |
| Weak or ambiguous growth | Adjust selection stringency and confirm hits by point-to-point testing. |
Sample and Information Requirements
- Bait gene or protein sequence
- Known domains and predicted activation regions, if available
- Plant species and sample source for prey library
- Existing plasmid or codon-optimized sequence, if available
- Number of bait constructs
- Desired validation and reporting format
Related Services
Please contact us with your bait protein sequence and screening objective for nuclear Y2H project evaluation.
For research use only, not for clinical use.