The CheckMate/Flexi Vector Mammalian-Two Hybrid System applies the principles of the yeast two-hybrid system to mammalian systems. Putative interacting proteins are inserted into the BIND (pFN11A) and ACT (pFN10A) vectors and transfected with the reporter construct, pGL4.31. If the proteins interact, they bring the GAL4 DNA Binding domain (BIND) in close proximity to the VP16 activation domain (ACT) initiating transcription on the pGL4.31 reporter construct, leading to generation of the firefly luciferase. Successful interactions can be assessed with the Dual-Luciferase Reporter Assay System. The pFN11A vector also serves as the control with SV40-promoter-driven Renilla luciferase production.|
Improved Sensitivity and Biological Relevance Due to: Increased Reporter Gene Expression: Codon optimization of synthetic genes for mammalian expression.Reduced Background and Risk of Expression Artifacts: Removal of cryptic DNA regulatory elements and transcription factor binding sites.Improved Temporal Response: Rapid Response™ technology available using destabilized luciferase genes.Additional Advantages Include:Flexible Detection Options: Choice of either synthetic luc2 (Photinus pyralis) or hRluc (Renilla reniformis) reporter genes.Easy Transition from Transient to Stable Cells: Choice of mammalian selectable markers.Easy Transfer from Vector to Vector: Common multiple cloning site and a unique SfiI transfer scheme.