Engineering single Cys2His2 zinc finger domains using a bacterial cell-based two-hybrid selection system.
Individual synthetic Cys2His2 zinc finger domains with novel DNA-binding specificities can be identified from large randomized libraries using selection methodologies such as phage display. We have previously demonstrated that a bacterial cell-based two-hybrid system is at least as effective as phage display for selecting zinc fingers with desired specificities from such libraries. In this chapter we provide updated, detailed protocols for performing zinc finger selections using the bacterial two-hybrid system.
Thibodeau-Beganny
Stacey
S
Molecular Pathology Unit, Center for Cancer Research, Massachusetts General Hospital, Charlestown, MA, USA.
Maeder
Morgan L
ML
Joung
J Keith
JK
D001419:Bacteria / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D008954:Models, Biological; D015202:Protein Engineering / Q000379:methods*; D011995:Recombination, Genetic / Q000235:genetics; D020798:Two-Hybrid System Techniques; D016335:Zinc Fingers / Q000235:genetics*
D004268:DNA-Binding Proteins