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300 Serebriiskii et al.Table 2PlasmidsShort name/Antibioticsource Full name marker Comment/descriptionpBaitC a pGLS23 a Cm R Basic plasmid to clone bait asa fusion to λcI proteinpTRG b pTRG Tc R Basic plasmid to clone preyas fusion to E. coli RNAPα-subunit residues 1–248pLibB a pBR-UV5-αLP Ap R Basic plasmid to clone prey asa fusion to E. coli RNAPα-subunit residues 1–248;has f1 origin of replicationpTRG-RGL pTRG-RGL-2 Tc R Control plasmid that expresses(control)an activation domain-RGLfusion proteinpBaitC-Ras a pGLS22-HRas Cm R Bait plasmid expresses a λcI-(control)HRasfusion; positive controlfor interaction with BRaf andRGL-2pBaitC-zip a pGLS22- Cm R Bait plasmid that expresses a(control) EE 12345λcI-leucine zipper proteinfusion; negative control forinteractionpLibB-Raf a pBR-UV5- Ap R Control plasmid that expresses(control) αLP-BRaf an activation domain-BRaffusionExpression of the fusion proteins in E. coli is driven by tandem lpp/lacUV5 promoters.All plasmids except for pAC-UV5-αLP have a pBR322 E. coli replication origin.It is noted, pGLS23 is described as the prototypical cI-fusion (pBaitC) plasmid: this dual-hostexpression vector also allows the screening in a Dual Bait yeast two-hybrid system, as describedin Tikhmyanova et al. Chapter 15.a Golemis lab (Fox Chase Cancer Center, Philadelphia, PA).b Stratagene.2.4. VectorsFigure 3 and Table 2 summarize and provide maps and other information forthe bacterial two-hybrid vectors used in this method (see Note 1).2.5. Bacterial StrainsSee Table 3 for genotypes of the E. coli strains used for selection.