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PMID: 18314590 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Engineering Cys2His2 zinc finger domains using a bacterial cell-based two-hybrid selection system.

Methods in molecular biology (Clifton, N.J.) ·Vol. 408 ·2007-00-00 ·Pages 317-34

Thibodeau-Beganny S, Joung JK

Abstract

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.

MeSH Terms
Amino Acid Sequence Bacteria/genetics Base Sequence DNA Primers/genetics F Factor/genetics Genes, Reporter Molecular Sequence Data Peptide Library Plasmids/genetics Protein Engineering/methods Recombination, Genetic Two-Hybrid System Techniques Zinc Fingers/genetics
Chemicals
DNA Primers Peptide Library
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Thibodeau-Beganny Stacey
Massachusetts General Hospital, Charlestown, USA.
Joung J Keith
Article Info
Journal
Methods in molecular biology (Clifton, N.J.)
Abbr.
Methods Mol Biol
ISSN
1064-3745
Published
2007-00-00
Pages
317-34
Language
English
Region
United States
NLM ID
9214969
Subset
IM
Grants
NIH HHS · DP1 OD006862 · United States
NIDDK NIH HHS · K08 DK002883 · United States
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