Cloning and expression in Escherichia coli of the gene encoding the [2Fe-2S] ferredoxin from Clostridium pasteurianum.
The gene encoding the [2Fe-2S] ferredoxin from Clostridium pasteurianum has been amplified from genomic DNA by the polymerase chain reaction and cloned under the control of a promoter specifically recognized by T7 RNA polymerase. The protein has been overproduced in E. coli and found to be identical to its native counterpart purified from C. pasteurianum, including the molecular weight, the N-terminal sequence and the spectroscopic properties of the [2Fe-2S] chromophore.
Fujinaga
J
J
Département de Biologie Moléculaire et Structurale, Grenoble, France.
Meyer
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular / Q000379:methods; D003013:Clostridium / Q000235:genetics* / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005288:Ferredoxins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D015183:Restriction Mapping
D004269:DNA, Bacterial; D005288:Ferredoxins; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins