Cloning and expression of the ferredoxin gene from extremely halophilic archaeon Haloarculajaponica strain TR-1.
The gene encoding a ferredoxin (Fd) from Haloarculajaponica strain TR-1 was cloned and sequenced. Sequence analysis of the cloned Ha. japonica Fd gene revealed that the structural gene consisted of an open reading frame of 387 nucleotides encoding 129 amino acids. The deduced amino acid sequence of Ha. japonica Fd showed 84 to 98% identity with corresponding sequences in other extremely halophilic archaea. The Ha. japonica Fd gene was inserted into the shuttle vector pWL102 and used to transform Ha. japonica. Ha. japonica Fd could then be produced as a fusion with His-Tag (6xHis) in Ha. japonica host cells. The absorption and ESR spectra of the Fd/His Tag fusion protein revealed the presence of a [2Fe-2S] cluster which is characteristic of native Ha. japonica Fd.
Matsuo
T
T
Department of Bioengineering, Tokyo Institute of Technology, Yokohama, Japan.
Ikeda
A
A
Seki
H
H
Ichimata
T
T
Sugimori
D
D
Nakamura
S
S
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004578:Electron Spin Resonance Spectroscopy; D005288:Ferredoxins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D019613:Haloarcula / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment
D019843:Archaeal Proteins; D005288:Ferredoxins; D011993:Recombinant Fusion Proteins