© 1995 • 280 pp., illus., appendices, index (volume 1 of a three-book set)
Plastic comb binding • ISBN  978-087969438-8

Foreword by Carl R. Woese, University of Illinois

The use of ribosomal RNA sequences as molecular chronometers has led to the recognition of a group of prokaryotes, the Archaea, that are phylogenetically distinct from both bacteria and eukaryotes. Although the Archaea are prokaryotes, lacking a nuclear membrane and possessing a single circular chromosome, they possess several molecular properties with similarity to the eukaryotes such as transcription signals, transcription factors, chaperones, and histones. Their unique phylogeny is underscored by a predilection for extreme environments which include high temperature, high salt, and strictly anaerobic conditions and the adaptive strategies that they have evolved to survive in these environments. These unique biomolecular adaptations have recently generated interest in the areas of genetic variability and genome evolution, extremely thermostable enzymes, such as DNA polymerases for PCR, methanogenesis as an alternative fuel source, anaerobic bioremediation, and use of bacteriorhodopsin for production of a biocomputer chip or a model for transmembrane ion translocation.

The Archaea occupy a pivotal phylogenetic position and offer extraordinary potential for biotechnology. One of the difficulties facing researchers, especially newcomers who wish to study the Archaea, has been the lack of a comprehensive source for the specialized techniques required. As the field has evolved, solutions to many problems have been discovered empirically and the special techniques have largely been disseminated either verbally or by means of journal articles. The protocols in these three books are selected to provide a detailed guide to experiments with the methanogenic, extremely halophilic, and thermophilic sulfur-utilizing Archaea, with overviews to highlight areas of future development. The individual protocols consist of an introduction describing the specific applications of the techniques, step-by-step procedures for applying the protocols, followed by any additional comments that will facilitate successful application of the protocol. A feature of research in this area is the interplay between microbiology, bioengineering, bio-chemistry, and molecular biology, and authors from all of these fields have been selected to provide these three concise and comprehensive resources for scientists interested in conducting research on the Archaea.

• Volume 1: Halophilic Archaea
• Volume 2: Methanogenic Archaea
• Volume 3: Thermophilic Archaea

Halophilic Archaea: An Overview (S. DasSarma)

Growth and Identification: Protocol 1. Cultivation of Halophilic Archaea (F. Rodriguez-Valera); Protocol 2. Growth of Halophilic Archaea Under Conditions of Low Oxygen Tension and High Light Intensity (M.C. Betlach, R.F. Shand); Protocol 3. Selection and Screening Methods for Halobacterial Rhodopsin Mutants (J.L. Spudich, E.N. Spudich); Protocol 4. Electron Microscopy of Halobacteria (Y. Nishiyama et al.)

Biochemistry: Protocol 5. Isoprenoids and Polar Lipids of Extreme Halophiles (M. Kates, S. Kushwaha); Protocol 6. Isolation of Purple Membranes (D. Oesterhelt); Protocol 7. Purification of Halorhodopsin from Halobacterium halobium (J.K. Lanyi); Protocol 8. Spectroscopic Assays for Sensory Rhodopsins I and II in Halobacterium salinarium Cells and Enriched Membrane Preparations (R.A. Bogomolni et al.); Protocol 9. Purification of the ATPase from Halobacterium halobium (B. Schobert); Protocol 10. Electrophoresis of Halobacterium Gas Vesicles in Phenol-Acetic Acid-Urea Gels (R.D. Simon); Protocol 11. Isolation of Halobacterial Ferredoxin (D. Oesterhelt, L. Kerscher); Protocol 12. Halophilic Proteases from Halophilic Archaea (M. Kamekura); Protocol 13. Purification of Halophilic Archaeal Ketohexokinase from Haloarcula vallismortis (V. Rangaswamy, W. Altekar); Protocol 14. Purification of Superoxide Dismutase and a Catalase-Peroxidase in a High-salt Environment (M.L. Salin, N.J. Brown-Peterson);

Molecular Biology and Genetics: Protocol 15. Purification of Halobacterium halobium DNA Polymerases (M. Kohiyama et al.); Protocol 16. Purification of DNA-dependent RNA Polymerase from Halobacterium halobium (F. Gropp, J. Leibrock); Protocol 17. Preparation of Transfer RNA, Aminoacyl-tRNA Synthetases, and tRNAs Specific for an Amino Acid from Extreme Halophiles (R. Gupta); Protocol 18. Cell-free Protein Synthesis System for Halobacterial Archaea (I. Marin et al.); Protocol 19. Isolation of Free and Membrane-bound Polysomes from Halobacterium halobium (R. Gropp); Protocol 20. Total Reconstitution of Ribosomal Subunits of Haloferax mediterranei (M.E. Sanchez, R. Amils); Protocol 21. The MPD-NaClH2O System for the Crystalization of Halophilic Proteins (S. Richard et al.); Protocol 22. Purification and Labeling of RNA from Halobacterium halobium for Identification of Regulated Genes (S. DasSarma, J.-M. Kim); Protocol 23. A Rapid Procedure for the Isolation of RNA from Haloferax volcanii (D.T. Nieuwlandt et al.); Protocol 24. Analysis of RNA Transcripts in Archaea (P.P. Dennis, J. Chow); Protocol 25. Isolation of Genomic and Plasmid DNAs from Halobacterium halobium (W.-L. Ng et al.); Protocol 26. Preparation of Intact, Agarose-embedded DNA from Halobacterium halobium and Its Digestion by Restriction Enzyme (N. Hackett); Protocol 27. Fractionation of Halobacterial DNA into FI and FII Using Affinity Chromatography on Malachite Green Bisacrylamide (F. Pfeifer); Protocol 28. Electrophoretic Mobility Shift Assays with Crude Extracts of Halobacterium halobium (N. Hackett, R. Ken); Protocol 29. Transformation of Halophilic Archaea (S.W. Cline et al.); Protocol 30. Gene Replacement in Halobacterium halobium (M. Krebs); Protocol 31. Selection for Spontaneous and Engineered Mutations in the rRNA Genes in Halophilic Archaea (A.S. Mankin)

Appendices