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Cloning, sequencing and overexpression of the Desulfovibrio gigas ferredoxin gene in E. coli
Author(s) -
Chen Baowei,
Me Nanda K.,
Dervertarnian Lisa,
Moura Jose J.G.,
Przybyla Alan E.
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00891-4
Subject(s) - cloning (programming) , ferredoxin , gene , microbiology and biotechnology , biology , desulfovibrio , molecular cloning , genetics , escherichia coli , bacteria , biochemistry , peptide sequence , enzyme , computer science , programming language
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.