Engineering Cys2His2 zinc finger domains using a bacterial cell-based two-hybrid selection system.
Synthetic Cys2His2 zinc finger domains with novel DNA-binding specificities can be identified from large randomized libraries using selection methodologies such as phage display. It has been 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 these libraries. In this chapter the authors provide updated and detailed protocols for performing zinc finger selections using the bacterial two-hybrid system.
Thibodeau-Beganny
Stacey
S
Massachusetts General Hospital, Charlestown, USA.
Joung
J Keith
JK
D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005144:F Factor / Q000235:genetics; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011995:Recombination, Genetic; D020798:Two-Hybrid System Techniques; D016335:Zinc Fingers / Q000235:genetics*
D017931:DNA Primers; D019151:Peptide Library