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Nuclear Protein Y2H cDNA Library Screening

Protein-protein interactions control variety of biological phenomena including development, cell to cell interactions and metabolic processes. Thus the knowledge of an organism's protein interaction network provides insights into the functions of individual proteins, the structure of functional complexes, and eventually, the organization of the whole cell. With the development of biotechnology, the protein-protein interactions can be detected by plenty of experimental methods. However, among these methods, the yeast two-hybrid system (Y2H) is the most popular method to identify protein interaction in vivo.

Schematic of nuclear protein Y2H screening. Figure 1. Schematic of nuclear protein Y2H screening.[1]

Two-hybrid screening (originally known as yeast two-hybrid system or Y2H) was originally devised by Fields and Song as a protein interaction detection system in yeast. Subsequently, it has been employed in many laboratories as a means of screening cDNA and genomic fusion libraries for protein interaction partners. Creative Biogene's nuclear protein Y2H library screening service is developed base on this technology which can identify proteins that interact with a protein of your interest using your pre-made library or our custom-built Y2H library. In our service, a bait protein of your interest is fused to a Gal4 DNA-binding domain and the library of proteins of interest 'ORF' (prey) is fused to Gal4 transcriptional activation domain (AD). Cotransform the two plasmids expressing bait and prey fusion proteins in the same cell. If, proteins bait and prey interact in the cells, they reconstitute a transcription factor which activates a reporter gene (eg. HIS3) and therefore allows the cell to grow on selective synthetic media (media lacking histidine). Thus, we can finally obtain the 'ORFs' that interact with your interest protein.

Applications:

  • Discover novel protein partners for your favorite protein
  • Elucidate mechanisms of action
  • Ascribe functions to uncharacterized proteins
  • Unravel pathways and molecular mechanism

As a leading biotechnology company in the world, Creative Biogene has extensive expertise and experience to provide you with nuclear protein Y2H library screening service. Creative Biogene possesses sophisticated equipment, advanced technologies, highly experienced staffs which are able to provide you personalized screening services to meet your analysis needs. Please do not hesitate to contact us at or 1-631-626-9181. We are looking forward to working together with your attractive projects.

References:

  1. Xing S., et al. (2016). Techniques for the analysis of protein-protein interactions in vivo. Plant physiology, 171(2), 727-758.
  2. Zhu L. (1997). Yeast GAL4 two-hybrid system. InRecombinant Protein Protocols (pp. 173-196). Humana Press.
  3. Cowell IG. (1997). Yeast two-hybrid library screening. IncDNA Library Protocols (pp. 185-202). Humana Press.
For research use only. Not intended for any clinical use.

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