Yeast Two-hybrid (Nuclear System) Screening

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Amerigo Scientific offers advanced Yeast two-hybrid (Y2H) screening service for the comprehensive analysis of protein-protein interaction using extensively validated nuclear protein cDNA libraries. Coverage of over 100 species and expert bioinformatics support ensure high-quality results for research needs.

The yeast two-hybrid system is the primary method for studying protein interactions in living cells. The classical Gal4 transcription factor system is utilized, featuring a DNA-binding domain (BD) and a transcriptional activation domain (AD). When target proteins are fused to these domains, their interaction restores transcriptional activity, enabling precise detection of the protein-protein interaction.

Figure 1. Technical Principle of Our Y2H (Nuclear System) Screening Service

Detailed Experimental Process

Figure 2. Experimental Process for Our Y2H (Nuclear System) Screening Service

Content of Our Y2H Screening Service

The total workflow of our Y2H screening service typically requires 65 working days.

Content Timeline Core Deliverables
Gene Synthesis and Codon Optimization 15 days
  • Validated positive clone sequences (≥15 sequences based on Sanger sequencing)
  • Bait plasmid (optional)
  • NGS sequencing results and bioinformatics analyses
  • Functional annotation (GO, KEGG) and interaction strength analysis
  • Detailed experimental report in Word format, including methodologies, step-by-step experimental details, high-resolution result images, and comprehensive data interpretation.
Autoactivation Testing 10 days
Yeast Transformation and Screening 20 days
Validation and Re-screening 5 days
NGS, Bioinformatics Analysis, and Reporting 15 days

Key Advantages of Our Y2H Screening Service

Enhanced Detection Accuracy

  • Dual-tier screening methodology
  • Three-deficiency media (SD-Trp/-Leu/-His)
  • Four-deficiency media (SD-Trp/-Leu/-His/-Ade)
  • Advanced X-α-gal assays

Dedicated Support Services

  • Expert consultation
  • Project-specific guidance
  • Ongoing technical support
  • Result interpretation assistance

Complete Documentation Package

  • Detailed experimental documentation
  • High-resolution result images
  • Comprehensive sequencing data (Sanger sequencing;  Next-generation sequencing (NGS))
  • Professional functional annotations

Advanced Bioinformatics Analysis

  • Professional data interpretation
  • Functional annotation
  • Transcription factor identification
  • Kinase analysis
  • Interaction strength quantification

Quality Assurance

  • Frame-shift analysis for positive clone verification
  • NGS-based high-throughput validation
  • Multiple quality control checkpoints

Nuclear Y2H One-to-one Validation Service

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Amerigo Scientific also offers a specialized Y2H one-to-one interaction validation service. This validation service is tailored for research that seeks to confirm specific protein-protein interactions and is used to validate screening results or test predefined interaction pairs.

Content Timeline Core Deliverables
Gene Synthesis and Vector Construction
  • Codon optimization and synthesis of target protein genes.
  • Construction of Protein A into the pGADT7 vector and Protein B into the pGBKT7 vector.
 10 days
  • Recombinant plasmids (Optional);
  • A digital experiment report, including methodologies, step-by-step experimental details, high-resolution result images, and comprehensive data interpretation.
Yeast Transformation and Autoactivation Testing
  • Transformation of recombinant plasmids into yeast cells.
  • Autoactivation testing using selective media to ensure experimental reliability.
 7 days
Interaction Validation
  • Setting up positive and negative controls for interaction verification.
  • Confirming protein-protein interactions using selective media and X-α-gal assays.
 10 days


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