© 1995 • 217 pp., illus., appendices, index (volume 3 of a three-book set)
Plastic comb binding • ISBN  0-87969-440-8

Foreword by Carl R. Woese, University of Illinois

The use of ribosomal RNA sequences as molecular chronometers hasled to the recognition of a group of prokaryotes, the Archaea, thatare phylogenetically distinct from both bacteria and eukaryotes.Although the Archaea are prokaryotes, lacking a nuclear membrane andpossessing a single circular chromosome, they possess severalmolecular properties with similarity to the eukaryotes such astranscription signals, transcription factors, chaperones, andhistones. Their unique phylogeny is underscored by a predilectionfor extreme environments which include high temperature, high salt,and strictly anaerobic conditions and the adaptive strategies thatthey have evolved to survive in these environments. These uniquebiomolecular adaptations have recently generated interest in theareas of genetic variability and genome evolution, extremelythermostable enzymes, such as DNA polymerases for PCR, methanogenesisas an alternative fuel source, anaerobic bioremediation, and use ofbacteriorhodopsin for production of a biocomputer chip or a model fortransmembrane ion translocation.

The Archaea occupy a pivotal phylogenetic position and offerextraordinary potential for biotechnology. One of the difficultiesfacing researchers, especially newcomers who wish to study theArchaea, has been the lack of a comprehensive source for thespecialized techniques required. As the field has evolved, solutionsto many problems have been discovered empirically and the specialtechniques have largely been disseminated either verbally or by meansof journal articles. The protocols in these three books are selectedto provide a detailed guide to experiments with themethanogenic, extremelyhalophilic, andthermophilic sulfur-utilizing Archaea,with overviews to highlight areas of future development. Theindividual protocols consist of an introduction describing thespecific applications of the techniques, step-by-step procedures forapplying the protocols, followed by any additional comments that willfacilitate successful application of the protocol. A feature ofresearch in this area is the interplay between microbiology,bioengineering, bio-chemistry, and molecular biology, and authorsfrom all of these fields have been selected to provide these threeconcise and comprehensive resources for scientists interested inconducting research on the Archaea.


• Volume 1: Halophilic Archaea

• Volume 2: Methanogenic Archaea

• Volume 3: ThermophilicArchaea


Thermophilic Archaea: An Overview (M.W.W. Adams)

Growth and Identification: Protocol 1. Isolation and Cultivation of Heterotrophic Hyperthermophiles from Deep-sea Hydrothermal Vents (H.W. Jannasch et al.); Protocol 2. Isolation, Growth, and Maintenance of Hyperthermophiles (J.A. Baross); Protocol 3. Plate Cultivation Technique for Strictly Anaerobic, Thermophilic, Sulfur-metabolizing Archaea (G. Erauso, D. Prieur); Protocol 4. Continuous Culture Techniques for Extremely Thermophilic and Hyperthermophilic Archaea (R.N. Schicho et al.); Protocol 5. High-pressure-High-temperature Cultivation of Extremely Thermophilic Archaea (C.M. Nelson, D.S. Clark); Protocol 6. Large-scale Growth of Hyperthermophiles (M.W.W. Adams); Protocol 7. Culture of Thermoplasma acidophilum (D. Searcy)

Biochemistry: Protocol 8. Solubilization from Membranes and Purification of Hydrogenases from Pyrodictium spp. (R.J. Maier et al.); Protocol 9. Purification of Hydrogenase and Ferredoxin from a Hyperthermophile (M.W.W. Adams, Z.H. Zhou); Protocol 10. Archaeal Lipid Analysis (D.B. Hedrick, D.C. White); Protocol 11. Glucose Metabolic Pathways in Thermoplasma acidophilum using 13C and 15N NMR Spectroscopy (K.J. Stevenson et al.)

Molecular Biology and Genetics: Protocol 12. Purification of Plasmids from Thermophilic and Hyperthermophilic Archaea (F. Charbonnier, P. Forterre); Protocol 13. Transfection of Sulfolobus solfataricus (C. Schleper, W. Zillig); Protocol 14. Preparation of Genomic DNA from Sulfur-dependent Hyperthermophilic Archaea (R. Vijayaragavan, M.W.W. Adams); Protocol 15. RNA Extraction from Sulfur-utilizing Thermophilic Archaea (J. DiRuggiero, F.T. Robb); Protocol 16. Reliable Amplification of Hyperthermophilic Archaeal 16S rRNA Genes by the Polymerase Chain Reaction (A.-L. Reysenbach, N.R. Pace); Protocol 17. Characterization of Archaeal Introns in the rRNA Genes of Hyperthermophiles (J.Z. Dalgaard et al.); Protocol 18. Generation of Subtraction Probes for Isolation of Specific Genes in Thermophilic Archaea (K.A. Robinson et al.); Protocol 19. Isolation of Sulfolobus acidocaldarius Mutants (D. Grogan); Protocol 20. Measurement of Mutation Rates in Sulfolobus acido-caldarius (D. Grogan); Protocol 21. Typing Marine Vent Thermophiles by DNA Polymerase Restriction Fragment Length Polymorphisms (F.B. Perler et al.); Protocol 22. Typing Hyperthermophilic Archaea Based on the 16S/23S rRNA Spacer Region (J. DiRuggiero et al.); Protocol 23. In Vitro Transcription from Natural and Mutated rDNA Promoters of the Extremely Thermophilic Archaeon Sulfolobus shibatae (J. Hain, W. Zillig)

Appendices