DUALhybrid kit

DUALhybrid technology enables you to screen a protein of interest for interactors expressed from a cDNA library of interest. Two different bait vectors give you the option to express your protein of interest either as N- or C-terminal fusion to the LexA domain. The choice of two orientations enhances your chances of finding all interactors for your protein and reduces the chances of self-activation.

The triple reporter strain NMY51 allows high stringency selection and greatly lowers the rate of false positives in a screen.

The DUALhybrid technology

The bait

A bait is constructed by fusing a protein of interest to the LexA domain. The LexA domain localizes the bait to a particular DNA sequence upstream of a reporter gene.

The prey

A prey is constructed by fusing a second protein of interest to the activation domain of a transcription factor.

Interaction between bait and prey

An interaction between the two proteins recruits the prey to the start of the reporter gene and the hybrid transcription factor assembled in this way from bait and prey (hence “two-hybrid”) activates the reporter gene. A protein interaction is thus measured by reporter gene activity. Commonly used reporter genes are either auxotrophic markers which allow selection on a minimal medium or color reporters like lacZ.

Manual -DUALhybrid kit

DUALhybrid vector selection guide

MSDS - DUALhybrid cDNA libraries

MSDS - DUALhybrid cDNA libraries

MSDS - NMY51 yeast reporter strain

Literature

A novel genetic system to detect protein-protein interactions.
Fields and Song Nature. 340, 245-6 (1989)
LexA-based two-hybrid systems.
Fashena et al. Methods Enzymol. 328, 14-26 (2000)

Publications

SLITRK1 binds 14-3-3 and regulates neurite outgrowth in a phosphorylation-dependent manner
Kajiwara et al. Biol Psychiatry. Nov 15;66(10):918-25. (2009)

FE65 Binds Teashirt, Inhibiting Expression of the Primate-Specific Caspase-4
Kajiwara et al.  PLoS ONE. 4(4) (2009): e5071.

Molecular dissection of NRG1-ERBB4 signaling implicates PTPRZ1 as a potential schizophrenia susceptibility gene.
Buxbaum et al.  Mol. Psychiatry. Apr 17 (2007) [Epub ahead of print]

XLF Interacts with the XRCC4-DNA Ligase IV Complex to Promote DNA Nonhomologous End-Joining
Ahnesorg et al. Cell 124(2), 301-313 (2006)

Promyelocytic leukemia zinc finger protein localizes to the cochlear outer hair cells and interacts with prestin, the outer hair cell motor protein
Nagy et al. Hearing Research 204, 216-222 (2005)

NEDD8 Ultimate Buster-1L Interacts with the Ubiquitin-like Protein FAT10 and Accelerates Its Degradation. (off site, PDF)
Hipp et al. J. Biol. Chem. 279 (16), 16503-16510 (2004)

Brain-type creatine kinase BB-CK interacts with the Golgi Matrix Protein GM130 in early prophase
Burklen et al. Mol. Cell. Biochem. 297, 53-64 (2007)