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. 1994 Aug;176(15):4790–4793. doi: 10.1128/jb.176.15.4790-4793.1994

Cloning, expression, and molecular characterization of the gene encoding an extremely thermostable [4Fe-4S] ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus.

A Heltzel 1, E T Smith 1, Z H Zhou 1, J M Blamey 1, M W Adams 1
PMCID: PMC196307  PMID: 8045914

Abstract

The gene for ferredoxin from the hyperthermophilic archaeon Pyrococcus furiosus was cloned, sequenced, and expressed in Escherichia coli. The coding region confirmed the determined amino acid sequence. Putative archaeon-type transcriptional regulatory elements were identified. The fdxA gene appears to be an independent transcriptional unit. Recombinant ferredoxin was indistinguishable from the protein purified from P. furiosus in its thermal stability and in the potentiometric and spectroscopic properties of its [4Fe-4S] cluster.

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Selected References

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  1. Adams M. W. Enzymes and proteins from organisms that grow near and above 100 degrees C. Annu Rev Microbiol. 1993;47:627–658. doi: 10.1146/annurev.mi.47.100193.003211. [DOI] [PubMed] [Google Scholar]
  2. Alting-Mees M. A., Short J. M. pBluescript II: gene mapping vectors. Nucleic Acids Res. 1989 Nov 25;17(22):9494–9494. doi: 10.1093/nar/17.22.9494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Amann E., Ochs B., Abel K. J. Tightly regulated tac promoter vectors useful for the expression of unfused and fused proteins in Escherichia coli. Gene. 1988 Sep 30;69(2):301–315. doi: 10.1016/0378-1119(88)90440-4. [DOI] [PubMed] [Google Scholar]
  4. Aono S., Bryant F. O., Adams M. W. A novel and remarkably thermostable ferredoxin from the hyperthermophilic archaebacterium Pyrococcus furiosus. J Bacteriol. 1989 Jun;171(6):3433–3439. doi: 10.1128/jb.171.6.3433-3439.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blamey J. M., Adams M. W. Purification and characterization of pyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeon Pyrococcus furiosus. Biochim Biophys Acta. 1993 Jan 15;1161(1):19–27. doi: 10.1016/0167-4838(93)90190-3. [DOI] [PubMed] [Google Scholar]
  6. Brown J. W., Daniels C. J., Reeve J. N. Gene structure, organization, and expression in archaebacteria. Crit Rev Microbiol. 1989;16(4):287–338. doi: 10.3109/10408418909105479. [DOI] [PubMed] [Google Scholar]
  7. Bryant F. O., Adams M. W. Characterization of hydrogenase from the hyperthermophilic archaebacterium, Pyrococcus furiosus. J Biol Chem. 1989 Mar 25;264(9):5070–5079. [PubMed] [Google Scholar]
  8. Busse S. C., La Mar G. N., Yu L. P., Howard J. B., Smith E. T., Zhou Z. H., Adams M. W. Proton NMR investigation of the oxidized three-iron clusters in the ferredoxins from the hyperthermophilic archae Pyrococcus furiosus and Thermococcus litoralis. Biochemistry. 1992 Dec 1;31(47):11952–11962. doi: 10.1021/bi00162a038. [DOI] [PubMed] [Google Scholar]
  9. Conover R. C., Kowal A. T., Fu W. G., Park J. B., Aono S., Adams M. W., Johnson M. K. Spectroscopic characterization of the novel iron-sulfur cluster in Pyrococcus furiosus ferredoxin. J Biol Chem. 1990 May 25;265(15):8533–8541. [PubMed] [Google Scholar]
  10. DiRuggiero J., Robb F. T., Jagus R., Klump H. H., Borges K. M., Kessel M., Mai X., Adams M. W. Characterization, cloning, and in vitro expression of the extremely thermostable glutamate dehydrogenase from the hyperthermophilic Archaeon, ES4. J Biol Chem. 1993 Aug 25;268(24):17767–17774. [PubMed] [Google Scholar]
  11. Hain J., Reiter W. D., Hüdepohl U., Zillig W. Elements of an archaeal promoter defined by mutational analysis. Nucleic Acids Res. 1992 Oct 25;20(20):5423–5428. doi: 10.1093/nar/20.20.5423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laderman K. A., Asada K., Uemori T., Mukai H., Taguchi Y., Kato I., Anfinsen C. B. Alpha-amylase from the hyperthermophilic archaebacterium Pyrococcus furiosus. Cloning and sequencing of the gene and expression in Escherichia coli. J Biol Chem. 1993 Nov 15;268(32):24402–24407. [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Reiter W. D., Hüdepohl U., Zillig W. Mutational analysis of an archaebacterial promoter: essential role of a TATA box for transcription efficiency and start-site selection in vitro. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9509–9513. doi: 10.1073/pnas.87.24.9509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Reiter W. D., Palm P., Zillig W. Analysis of transcription in the archaebacterium Sulfolobus indicates that archaebacterial promoters are homologous to eukaryotic pol II promoters. Nucleic Acids Res. 1988 Jan 11;16(1):1–19. doi: 10.1093/nar/16.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Reiter W. D., Palm P., Zillig W. Transcription termination in the archaebacterium Sulfolobus: signal structures and linkage to transcription initiation. Nucleic Acids Res. 1988 Mar 25;16(6):2445–2459. doi: 10.1093/nar/16.6.2445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  18. Zwickl P., Fabry S., Bogedain C., Haas A., Hensel R. Glyceraldehyde-3-phosphate dehydrogenase from the hyperthermophilic archaebacterium Pyrococcus woesei: characterization of the enzyme, cloning and sequencing of the gene, and expression in Escherichia coli. J Bacteriol. 1990 Aug;172(8):4329–4338. doi: 10.1128/jb.172.8.4329-4338.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

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