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What is the DupLEX-ATM Yeast Two-Hybrid System? The DupLEX-ATM Yeast Two-Hybrid System is a LexA-based version of the yeast two-hybrid system using licensed technology.* The DupLEX-ATM Yeast Two-Hybrid System is conceptually the same as the Gal4-based system but is more versatile and contains many more useful features than the Gal4-based system. The DupLEX-ATM system uses LexA as the DNA binding protein and B42 as the transcriptional activation domain. Both are bacterial proteins, thus reducing the risk of endogenous yeast proteins binding to one of the two proteins and possibly creating a false positive interaction. [Note: The bacterial LexA protein does not repress transcription in yeast.] In addition, since Gal4p is not used as a component of the interactors in the DupLEX-ATM system, it is free to be used as an inducible regulator of target gene expression. Advantages of DupLEX-ATM over other systems fast cloning of genes for potential interacting proteins inducible expression of library proteins allows toxic interactors to be found reporter genes of differing sensitivities can allow autoactivating proteins to be screened many control and test plasmids are included to save time and effort libraries (available separately) are provided not only as primary transformant cells but also as ready-to use DNA purified directly from the primary transformants (not re-amplified) LexA and B42 activation proteins are heterologous to yeast, reducing the chances of obtaining false positives in the screen Is that all? No, that's not all. The DupLEX-ATM system also contains tests for determining if your bait protein can enter the yeast nucleus when fused to LexA and, if it can, to determine if it's able to bind to LexA DNA binding sites in vivo. LexA fusion proteins are normally translocated to the nucleus in yeast; however, if the test shows that your bait fusion is not getting into the nucleus, our system provides a bait fusion vector containing an SV40 nuclear localization signal. These controls, along with the testing of reporters of varying sensitivities, will greatly increase your chances of performing a successful screen and may save you much time and effort. Do I have to make my own library? Of course not! We take the work out of library preparation by supplying you with ready-to-use plasmid library DNA, in addition to primary transformant cells. The DNA is purified from the primary bacterial transformants. There is no need for tedious library titering or the potential loss of rare clones through re-amplification of the library. In addition, each library meets or exceeds our strict minimum standards and quality control criteria (ask for details). We have a number of libraries currently available and will be introducing more in the near future. If you have a particular library that you would like made and we don't have it, give us a call! We're here for you!   pGilda, a plasmid used for examining potentially toxic baits, is now available from OriGene Technologies!

Item Description Catalog# DupLEX-A System kit contains all items listed below DKT-100 Carrier DNA sonicated salmon sperm DNA, 5mg/ml DSS-100 5´ bait primer for sequencing bait-fusion protein junction DPR-100 5´ target primer for sequencing and PCR DPR-102 3´ target primer for sequencing and PCR DPR-103 Strains EGY48 (yeast) contains most sensitive LEU2 reporter DYS-48 EGY194 (yeast) contains medium sensitivity LEU2 reporter DYS-194 EGY188 (yeast) contains least sensitive LEU2 reporter DYS-188 EGY40 (yeast) contains no LEU2 reporter DYS-40 RFY206 (yeast) used in mating assay (controls) DYS-206 KC8 (E. coli) trp- strain used for rescuing target plasmids from yeast DES-108 Plasmids pEG202 used for making LexA-bait fusion protein DPL-100 pJK202 like pEG202, but contains nuclear localization sequence fused to LexA DPL-101 pJG4-5 used for making B42-cDNA libraries DPL-102 pJG4-6 similar to pJG4-5 but lacks the B42 domain DPL-103 pSH18-34 contains most sensitive lacZ reporter DPL-104 pJK103 contains medium sensitivity lacZ reporter DPL-105 pRB1840 contains least sensitive lacZ reporter DPL-106 pJK101 used to test bait translocation to nucleus and LexA operator binding DPL-107 pSH17-4 expresses LexA-Gal4p activation domain; positive control for reporters DPL-108 pRHFM1 expresses LexA-Drosophila bicoid gene (negative control for target fusions) DPL-109 pLexA-Max expresses LexA-Max (negative control for target fusions) DPL-110 pNLexA used for fusing LexA to the C-terminus rather than the N-terminus of the bait pBAIT DPL-111 pBAIT LexA-bait fusion that interacts with pTARGET DPL-112 pTARGET B42-target fusion that interacts with pBAIT DPL-113 NOTE: All components are subjected to our strict quality control standards before shipping. cDNA libraries may not be amplified, recombined, and/or resold without express written permission from OriGene Technologies, Inc. *Gyuris, J., Golemis, E.A., Chertkov, H. and Brent, R. (1993) "Cdil, a human G1- and S- phase protein phosphatase that associates with Cdk2", Cell 75: 791-803.