DUALhunter System

DUALhunter Kit (Cat # P01005)

• Protein-Protein Interaction Screening
  
  
• Protein Expression
  
  
• Protein Extraction and Refolding
  
  
• Library Construction and Preparation
  
  
• Yeast Kits, Strains and Tools
  
  
• Anti-Yeast & DNA Repair Antibodies
  
  
• Custom Services

Due to their ease of use, genetic screening systems such as the yeast two-hybrid (y2h) system have been especially successful in identifying novel protein interactions. However, the y2h system is biased against certain classes of proteins, such as transcription factors. When fused to a DNA-binding domain, these proteins autonomously activate transcription because they contain sequence stretches that act as transcriptional activators for the RNA polymerase II complex. Since the output of the y2h system is based on transcriptional activation of a reporter gene, these proteins activate the reporter genes on their own, in the absence of a protein interaction. Such proteins are termed "self-activating" and they cannot be used in a classical yeast two-hybrid system. Unfortunately, many important proteins belong to this class, including transcription factors, regulatory proteins associated with DNA, checkpoint proteins like p53 or p65, and many other proteins.

With the DUALhunter kit you can screen full-length, transcriptionally active, soluble (cytosolic or nuclear) proteins against a cDNA library of your choice to identify novel interaction partners of your protein of interest.

• Quickly screen your protein of interest for novel interactors in vivo
• No need to truncate your protein of interest or to select subdomains
• Screen transcriptionally active proteins or self-activating proteins
• Easy subcloning of full-length cDNA inserts using Sfi I technology

Reporter strain with additional markers and improved stringency, resulting in fewer false positives

Protein interaction is detected outside of the nucleus and therefore works with all transcriptionally active proteins. In addition, it can also be used to screen any soluble protein, domain or protein fragment to uncover novel protein interactions. A protein of interest (the bait) is inserted between the membrane protein Ost4p and the C-terminal half of ubiquitin (Cub), followed by the artificial transcription factor LexA-VP16. A second protein (the prey) is fused to the mutated N-terminal half of ubiquitin (NubG). The detection of a protein interaction is based on the split-ubiquitin system. If bait and prey interact, Cub and NubG complement to form split-ubiquitin, followed by cleavage and translocation of LexA-VP16 to the nucleus and transcriptional activation of endogenous reporter genes. The protein interaction between bait and prey is detected using the output of the reporter genes, either via a growth selection on minimal medium, or via the color marker lacZ. Replacing the plasmid encoding the prey by a cDNA library expressing millions of different proteins or protein fragments allows the uncovery of novel protein interactions.

View Premade Libraries for the DUALhunter/DUALmembrane System