Yeast two-hybrid technology can reflect the interaction of different proteins in living cells. It has been widely used in the study of gene function because of its simplicity, sensitivity and high efficiency.
Figure 1. Yeast Two-Hybrid Assay to Identify Interacting Proteins (Paiano, 2019)
Creative BioMart can provide yeast two-hybrid experiment service for cytoplasmic protein, nuclear protein and membrane protein. We have rich experience in yeast library construction and library screening, which can help customers analyze and solve various problems encountered during experiments.
Figure 2. The classical yeast two-hybrid system (Brückner, 2009)
It is used to screen interacting proteins, identify/verify protein-protein interactions, explore protein-protein interaction mechanisms, and draw protein linkage maps.
Based on the traditional yeast two-hybrid system, the protein-protein interaction is screened using the separated ubiquitin system.
We provide library construction and library screening services
| Service project | Customers provide | Deliverables |
| Library construction | Tissues/Cells | Liquid of yeast library (titrated higher than 1 x 108 Pictures of PCR identification results of library (with minimum 90% positive rate) |
| Library screening | Protein sequence / gene sequence / plasmid | All original pictures in the screening process; Screening and sequencing results of all interacting prey proteins |
Library construction
Library screening
The results of yeast two-hybrid experiment can be further verified：
| Service project | Objective | Details | Timeline (weeks) |
| Co-IP | Verify the interaction between proteins | / | 20 |
| GST pull-down | Verify the interaction between proteins | GST pull-down+WB / silver staining | 20 |
Total RNA extraction is a key step in the whole project. Based on many years of library construction experience, Creative BioMart provides customers with 4 optimized total RNA extraction technology solutions, which can effectively ensure the purity and integrity of total RNA, and ensure the diversity of samples.
TRIZOL method for cells or animal tissues.
CTAB method for samples with ultra-low RNA content in plant samples.
SDS method for samples with high starch content.
Kit extraction for samples with high content of polysaccharides and polyphenols.
With our professional research and development team and the latest protein expression and purification resources, we promise to deliver high-quality products for our customers. Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.
References
1. Paiano A, Margiotta A, De Luca M, Bucci C. Yeast Two-Hybrid Assay to Identify Interacting Proteins. Curr Protoc Protein Sci. 2019 Feb;95(1):e70.
2. Brückner, A., Polge, C., Lentze, N., Auerbach, D., & Schlattner, U. (2009). Yeast two-hybrid, a powerful tool for systems biology. International journal of molecular sciences, 10(6), 2763–2788.
