Abstract
Ligand-dependent chimeric Cre recombinases are powerful tools to induce specific DNA rearrangements in cultured cells and in mice. We report here the construction and characterization of a series of chimeric recombinases, each consisting of Cre fused to a mutated human oestrogen receptor (ER) ligand-binding domain (LBD). Two new ligand-dependent recombinases which contain either the G400V/M543A/L544A or the G400V/L539A/L540A triple mutation of the human ER LBD are efficiently induced by the synthetic ER antagonists 4-hydroxytamoxifen (OHT) and ICI 182,780 (ICI), respectively, but are insensitive to 17 beta-oestradiol (E2). Both chimeric recombinases should be useful for efficient spatio-temporally controlled site-directed somatic mutagenesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Binding Sites
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Enzyme Induction / drug effects
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Estradiol / analogs & derivatives
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Estradiol / pharmacology
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Estrogen Antagonists / pharmacology*
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Fulvestrant
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Humans
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Integrases / biosynthesis
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Integrases / metabolism*
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Kinetics
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Mice
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Molecular Sequence Data
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Mutagenesis, Insertional
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Mutagenesis, Site-Directed
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Oligodeoxyribonucleotides
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Point Mutation
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Receptors, Estrogen / biosynthesis
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Receptors, Estrogen / chemistry
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Receptors, Estrogen / metabolism*
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Tamoxifen / analogs & derivatives
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Tamoxifen / pharmacology
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Teratoma
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Tumor Cells, Cultured
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Viral Proteins*
Substances
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Estrogen Antagonists
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Oligodeoxyribonucleotides
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Receptors, Estrogen
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Recombinant Fusion Proteins
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Viral Proteins
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Tamoxifen
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afimoxifene
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Fulvestrant
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Estradiol
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Cre recombinase
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Integrases