Yeast two-hybrid (Y2H) is a technique that uses genetics to analyze the interaction between proteins. In yeast, the DNA-binding domain (DNA-BD) and the activation domain (AD) alone cannot activate the transcription, and the two are sufficiently close in space to enable transcription of genes downstream of the promoter. Since the establishment of this method, after continuous improvement and development, it can not only detect the interaction between known proteins but also can be used to discover new unknown proteins that interact with known proteins.
Figure 1. Principle of the yeast two-hybrid system (Mehla, J.; et al. 2017)
The bacterial two-hybrid system (B2H) utilizes the characteristics that the calmodulin-dependent adenylate cyclase activation region of Bordetella pertussis can be divided into two relatively independent functional fragments, T18 and T25. When these two fragments are expressed independently in Escherichia coli BTH101 (cya-), they cannot recognize and interact with each other. These two functional regions are fused and expressed with a pair of interactable proteins, they can form a functional adenylate cyclase that complements the E. coli cya strain and increase the β-galactosidase activity of the strain.
Figure 2. Principle of the bacterial two-hybrid system (Mehla, J.; et al. 2017)
The current study of how proteins interact in cells is an important part of molecular biology. Since the advent of the two-hybrid system, it has been a powerful molecular tool. It is an indispensable in vivo research method for studying protein interactions and discovering unknown interacting protein molecules.
After more than ten years of technical summary, Creative Proteomics has developed a mature and reliable two-hybrid platform, which is composed of a large number of sophisticated experimental instruments and professional technicians. We provide time-saving one-stop services, including library construction to the final screening of positive clones, to ensure the transparency and high efficiency of the whole process. In addition, our two-hybrid platform has been improved and upgraded to significantly eliminate false positives and improve the sensitivity and specificity of the two-hybrid system.
Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.
Creative Proteomics is an international biotechnology company dedicated to the study of intermolecular interactions and other related fields. We have established and improved the two-hybrid technology platform, overcoming many shortcomings in this system to greatly improve the accuracy and specificity of the experiment. Our one-stop service aims to save time and money for customers.