Advanced Yeast One-Hybrid (Y1H) Service

Background Y1H Platform Service Portfolio Workflow Advantage FAQs Online Inquiry

For over two decades, Creative Biolabs has been at the forefront of molecular interaction analysis, offering a robust and sophisticated Yeast One-Hybrid (Y1H) screening platform to researchers worldwide. Leveraging our deep expertise and state-of-the-art technologies, we empower you to decipher complex gene regulatory networks. Our Y1H service is a powerful in vivo tool designed to efficiently screen vast cDNA libraries for proteins that bind to your specific DNA bait sequence, from known promoters and enhancers to novel regulatory motifs.

Discuss Your Project Today

Understanding the Yeast One-Hybrid (Y1H) System

The Yeast One-Hybrid (Y1H) system is a highly effective genetic method adapted from the renowned Yeast Two-Hybrid (Y2H) system. While Y2H identifies protein-protein interactions, Y1H is specifically engineered to detect and identify physical interactions between a protein (the "prey") and a known DNA sequence (the "bait").

The core principle relies on the modular nature of eukaryotic transcription factors, which typically have a DNA-binding domain (DBD) and a transcriptional activation domain (AD). In the Y1H system:

The Bait: Your DNA sequence of interest is cloned upstream of a reporter gene (e.g., HIS3, AUR1-C, LacZ) in a yeast expression vector. This creates a "bait-dependent" reporter strain.
The Prey: A cDNA or domain library is constructed where proteins are fused to a strong transcriptional activation domain (AD).
The Interaction: When the bait-reporter strain is transformed with the prey library, if a library-encoded protein (prey) binds to the bait DNA sequence, its fused AD is brought into proximity of the reporter gene's promoter, activating its transcription.
The Readout: This activation event allows the yeast cells to grow on selective media (e.g., medium lacking histidine or containing Aureobasidin A) or exhibit a colorimetric change, indicating a positive protein-DNA interaction.

Princple of modified Y1H system for transcription factors selection. (OA Literature). Fig. 1 Diagrammatic representation of the basis for the MEL1 selection strategy.¹

Our Advanced Y1H Technology Platform: Beyond the Standard Assay

At Creative Biolabs, we don't just offer a standard Y1H service; we provide a fully optimized, next-generation platform designed for maximum sensitivity and discovery potential.

Optimized Reporter Systems: We utilize multiple, highly stringent reporter genes, including nutritional markers (HIS3, URA3) and dominant selectable markers (AUR1-C), to minimize false positives and eliminate background growth.
High-Diversity Libraries: Success in Y1H hinges on the quality and complexity of the prey library. We offer a comprehensive collection of premade, high-diversity cDNA libraries from various species and tissues. We also specialize in custom cDNA library construction to meet the unique demands of your project.
Proprietary Vectors & Yeast Strains: Our proprietary vectors are engineered for high-level expression and stability, while our optimized yeast strains ensure robust reporter gene activation and low auto-activation, a common pitfall in Y1H screens.
Integration with NGS (Y1H-Seq): For complex screens yielding numerous hits, we can integrate Next-Generation Sequencing (NGS) to identify and quantify the abundance of interacting protein-encoding sequences. This provides a quantitative, genome-wide perspective on the "interactome" of your DNA bait.

Our Comprehensive Y1H Service Portfolio

We offer a flexible suite of services that can be used individually or as a complete package.

Y1H cDNA Library Screening Service: Our flagship service. You provide the DNA bait sequence, and we perform a full screen against a library of your choice to identify interacting proteins.
One-on-One Y1H Interaction Validation: If you have a putative protein-DNA pair, we can quickly and definitively validate the interaction using a focused Y1H assay.
Custom Bait and Vector Construction: We assist in designing and cloning your specific DNA bait sequence into our optimized Y1H reporter vectors.
Bait Auto-activation Testing: A crucial preliminary step. We rigorously test your bait sequence for any self-activation of the reporter gene to ensure the reliability of the subsequent screen.
Custom cDNA and gDNA Library Construction: If a suitable pre-made library is unavailable, our experts will construct a high-quality, project-specific library from your provided biological material.

Our Streamlined Y1H Screening Workflow

We follow a meticulous, step-by-step process to ensure transparency and high-quality outcomes.

Consultation: We begin with a deep dive into your research goals to design the optimal Y1H strategy.
Bait Construction: Your DNA sequence is cloned, and a bait-reporter yeast strain is generated and validated.
Auto-activation Test: We perform stringent tests to confirm the bait does not activate the reporters on its own.
Library Screening: The validated bait strain is transformed with a high-complexity prey library and plated on dual-selection medium.
Hit Identification: Positive colonies are isolated. The plasmids encoding the interacting "prey" proteins are recovered and their inserts are identified by DNA sequencing.
Reporting: You receive a detailed final report containing all methods, data, and a list of confirmed interacting partners.

The Creative Biolabs Advantage: Why Partner With Us?

Choosing Creative Biolabs for your Y1H screening project ensures you receive more than just data. You gain a dedicated research partner committed to your success.

Advantage	Description
Unrivaled Expertise	With over 20 years of experience, our scientists have mastered the nuances of the Y1H system, ensuring your project is designed and executed flawlessly.
Complete Customization	From bait design and library selection to screening stringency, we tailor every project parameter to your specific research goals.
High Success Rate	Our optimized platform and rigorous quality control, including a mandatory bait auto-activation test, lead to an industry-leading success rate.
High-Throughput Capability	Our automated platforms can screen millions of interactions rapidly, accelerating your research timeline without compromising on quality.
Comprehensive Deliverables	We provide a detailed report including raw data, hit sequence information, bioinformatics analysis, and full experimental details, ready for publication.
Transparent Communication	A dedicated Ph.D.-level project manager provides regular updates and is always available for consultation.

We offer a comprehensive, end-to-end Y3H service tailored to your specific project goals. Our expert team works closely with you from experimental design to final data analysis. Other optional protein-nucleic acid interaction (PNI) assay services:

Frequently Asked Questions (FAQs)

Q1: What kind of DNA sequence can be used as a Y1H bait?

A: Almost any DNA sequence between 20-500 bp can be used, including promoters, enhancers, silencers, or tandem repeats of a short motif. Very long sequences (>1 kb) may be challenging. We recommend providing a sequence that you hypothesize contains a protein-binding site.

Q2: What is bait auto-activation?

A: This occurs when the bait DNA sequence itself, without a binding partner, can weakly activate the reporter gene, leading to false positives. Our rigorous testing protocol identifies and mitigates this issue before the main screen begins.

Q3: What is the difference between Yeast One-Hybrid (Y1H) and ChIP-Seq?

A: Y1H identifies proteins that can bind to a known DNA sequence in vivo (in yeast). It answers: "What proteins bind to my DNA?". ChIP-Seq identifies the DNA sequences that a known protein does bind to in its native cellular context. It answers: "Where in the genome does my protein bind?". The two are highly complementary.

Q4: How many interacting proteins can I expect to find?

A: The number of hits can vary widely from zero to several dozen, depending on the nature of the bait sequence, its biological role, and the library being screened. Our goal is to provide you with a list of high-confidence interactors.

Embark on Your Discovery Journey Today

Unlock the secrets of your gene's regulatory landscape. With Creative Biolabs' expert Y1H services, you can confidently identify the key protein players governing your biological process of interest.

Contact Us Today for a Free Quote!

Reference

Lopato, Sergiy, et al. "Isolation of plant transcription factors using a modified yeast one-hybrid system." Plant methods 2 (2006): 1-15. Under Open Access license CC BY 4.0, without modification. https://doi.org/10.1186/1746-4811-2-3

×

Online Inquiry

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.