Plant BiFC Vector Construction and Fusion Design Service
BiFC Construct Design and Cloning
CD BioSciences provides plant BiFC vector construction and fusion design services to support interpretable protein-protein interaction imaging experiments.
The service focuses on sequence review, fusion orientation selection, reading-frame design, vector construction, clone screening, and sequence verification before plant transient expression and confocal imaging.
Why Fusion Design Matters
BiFC results can be affected by the position of the fluorescent fragment, protein folding, localization motifs, transmembrane regions, and interaction domains. In some projects, both N-terminal and C-terminal fusions should be considered to reduce false-negative risk and improve interpretability.
Figure 1. BiFC fusion orientation options for nYFP and cYFP constructs.
Service Content
Review of CDS, stop codon, reading frame, known domains, localization motifs, and construct feasibility.
Design of nYFP/cYFP, YFPn/YFPc, or related split-fluorescent protein fusions at suitable termini.
Gene synthesis, PCR amplification, restriction cloning, recombination cloning, or subcloning can be selected based on input material.
Positive colony screening, plasmid preparation, and Sanger sequencing can be included to confirm final constructs.
Construct QC Workflow
Figure 2. BiFC construct preparation and sequence verification workflow.
- Sequence and domain review
- Fusion orientation and vector design
- Primer or synthesis design
- Insert preparation and vector cloning
- Colony PCR or restriction screening
- Plasmid preparation
- Sanger sequencing or junction verification
- Construct summary for imaging work
Design Considerations
| Item | Why It Matters |
|---|---|
| N- or C-terminal tag position | Tag placement can affect protein folding, localization, and interaction-domain accessibility. |
| Stop codon strategy | Fusion constructs usually require stop codon handling to preserve the intended reading frame. |
| Linker design | A suitable linker can reduce steric hindrance between the protein and fluorescent fragment. |
| Localization or membrane domains | Signal peptides, transit peptides, and transmembrane regions may influence tag placement. |
| Control constructs | Empty-fragment and positive-control constructs are needed for result interpretation. |
Sample and Information Requirements
- Full CDS or protein sequences
- Species and gene identifiers
- Known domains or localization motifs
- Existing plasmids or templates, if available
- Vector map, resistance, and MCS information for supplied vectors
- Preferred fluorescent fragment system, if specified
- Required empty-vector or positive-control constructs
- Downstream imaging plan and expected plant expression system
Deliverables
- Fusion design summary
- Construct map or sequence information
- Colony screening records, when included
- Sanger sequencing or alignment results
- Verified plasmid DNA or glycerol stocks, when applicable
- Notes for downstream BiFC imaging setup
Related Services
Please contact us with the target sequences and preferred BiFC vector system for construct design review.
For research use only, not for clinical use.