Abstract
Most laboratory strains of the yeast Saccharomyces cerevisiae contain many copies of an autonomously replicating plasmid called 2-micron circle DNA. This plasmid codes for a site-specific recombinase, the FLP protein which promotes recombination across two 599-base pair inverted repeats of the plasmid DNA. We have cloned the FLP gene under the control of a strong Escherichia coli promoter and have hyperproduced the protein in that organism. Cell-free extracts from this source promote highly efficient site-specific recombination in vitro and we have used this activity to purify the FLP protein substantially. The enzyme acts efficiently on circular and linear substrates and requires only monovalent or divalent cations for activity.
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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Base Sequence
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DNA Nucleotidyltransferases / genetics*
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DNA Nucleotidyltransferases / isolation & purification
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DNA Restriction Enzymes
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Escherichia coli / genetics*
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Genes*
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Genes, Fungal*
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Plasmids*
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Saccharomyces cerevisiae / genetics*
Substances
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DNA Nucleotidyltransferases
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FLP recombinase
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DNA Restriction Enzymes
Associated data
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GENBANK/J01347
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GENBANK/L00321
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GENBANK/L00322
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GENBANK/L00323
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GENBANK/L00324
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GENBANK/M10185
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GENBANK/M11111
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GENBANK/M11593
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GENBANK/M14239
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GENBANK/M14240
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GENBANK/M14241
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GENBANK/M14242
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GENBANK/M14243
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GENBANK/M14244
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GENBANK/M14245
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GENBANK/M14253
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GENBANK/M14254
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GENBANK/M14255
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GENBANK/M14256
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GENBANK/M14257
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GENBANK/M14258
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GENBANK/M14259
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GENBANK/M14591
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GENBANK/M14592
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GENBANK/M14593
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GENBANK/M14594
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GENBANK/M14595
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GENBANK/M14596
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GENBANK/M14597
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GENBANK/M14598