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Isolation of novel single-chain Cro proteins targeted for binding to the bcl-2 transcription initiation site by repertoire selection and subunit combinatorics
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry, Biochemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry. Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Biochemistry and Organic Chemistry, Biochemistry.
2005 (English)In: Protein Engineering Design & Selection, ISSN 1741-0126, E-ISSN 1741-0134, Vol. 18, no 11, 537-546 p.Article in journal (Refereed) Published
Abstract [en]

New designed DNA-binding proteins may be recruited to act as transcriptional regulators and could provide new therapeutic agents in the treatment of genetic disorders such as cancer. We have isolated tailored DNA-binding proteins selected for affinity to a region spanning the transcription initiation site of the human bcl-2 gene. The proteins were derived from a single-chain derivative of the lambda Cro protein (scCro), randomly mutated in its recognition helices to construct libraries of protein variants of distinct DNA-binding properties. By phage display-afforded affinity selections combined with recombination of shuffled subunits, protein variants were isolated, which displayed high affinity for the target bcl-2 sequence, as determined by electrophoretic mobility shift and biosensor assays. The proteins analyzed were moderately sequence-specific but provide a starting point for further maturation of desired function.

Place, publisher, year, edition, pages
2005. Vol. 18, no 11, 537-546 p.
Keyword [en]
phage display, protein–DNA interactions, repressor, single-chain Cro, subunit shuffling
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-74639DOI: 10.1093/protein/gzi058PubMedID: 16186141OAI: oai:DiVA.org:uu-74639DiVA: diva2:102549
Available from: 2005-10-24 Created: 2005-10-24 Last updated: 2010-09-14Bibliographically approved

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Nilsson, Mikael T.I.Widersten, Mikael

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