Confirmation of the Fur operator site by insertion of a synthetic oligonucleotide into an operon fusion plasmid.
We constructed a synthetic oligonucleotide corresponding to the previously proposed consensus binding site for the Fur protein, a central iron-regulatory protein of Escherichia coli. When this oligonucleotide was introduced at the start of transcription of an operon fusion between the ompF promoter and the lacZ structural gene, beta-galactosidase activity became iron regulated. This consensus sequence is sufficient to function as an operator site for the binding of Fur protein in vivo.
Calderwood
S B
SB
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115.
Mekalanos
J J
JJ
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D007501:Iron / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides; D009875:Operator Regions, Genetic; D009876:Operon; D010957:Plasmids; D001616:beta-Galactosidase / Q000032:analysis
D001426:Bacterial Proteins; D009838:Oligodeoxyribonucleotides; D007501:Iron; D001616:beta-Galactosidase