Cloning, sequencing and overexpression of the Desulfovibrio gigas ferredoxin gene in E. coli.
We have cloned the gene encoding Desulfovibrio gigas ferredoxin using a photodigoxigenin-labelled probe synthesized with the polymerase chain reaction. The DNA sequence of the gene predicts a polypeptide of 58 residues after removal of the initial formyl methionine (polypeptide M(r) = 6,276). The ferredoxin gene was expressed in aerobically grown E. coli behind the lac promoter of pUC18 resulting in a high level of ferredoxin expression which comprises about 10% of the total cell protein. EPR analysis of recombinant ferredoxin revealed the presence of a [3Fe-4S] cluster which is characteristic of native D. gigas ferredoxin II.
Chen
B
B
Department of Biochemistry, University of Georgia, Athens 30602.
Menon
N K
NK
Dervertarnian
L
L
Moura
J J
JJ
Przybyla
A E
AE
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D003901:Desulfovibrio / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005288:Ferredoxins / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D013057:Spectrum Analysis
D004269:DNA, Bacterial; D005288:Ferredoxins; D011994:Recombinant Proteins