Description
Two-hybrid systems are extremely powerful methods for detecting protein:protein interactions in vivo. The basis of two-hybrid systems is the modular domains found in some transcription factors: a DNA-binding domain, which binds to a specific DNA sequence, and a transcriptional activation domain, which interacts with the basal transcriptional machinery. A transcriptional activation domain in association with a DNA-binding domain will promote the assembly of RNA polymerase II complexes at the TATA box and increase transcription.
In the CheckMate™ Mammalian Two-Hybrid System the DNA-binding domain and the transcriptional activation domain, produced by separate plasmids, are closely associated when one protein ("X") fused to a DNA-binding domain interacts with a second protein ("Y") fused to a transcriptional activation domain. In this system, interaction between proteins X and Y results in transcription of a reporter gene.
For more information, see the Protocols & Applications Guide .
Application
Confirming suspected interactions between two proteins and identifying domains/residues involved in protein:protein interactions.
Reference
Sadowski, I. et al. (1988) Nature 335 , 563–4.
pACT Vector GenBank® Accession Number AF264723 and vector sequence text file .
pBIND Vector GenBank® Accession Number AF264722 and vector sequence text file .
pG5luc Vector GenBank® Accession Number AF264724 and vector sequence text file .
