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Methods Mol Biol. 2011;765:435-45. doi: 10.1007/978-1-61779-197-0_26.

Engineering thermoacidophilic archaea using linear DNA recombination.

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School of Biological Sciences, University of Nebraska, Lincoln, NE, USA.


Thermoacidophilic archaea comprise one of the major classes of extremophiles. Most belong to the family Sulfolobales within the phylum Crenarchaeota. They are of applied interest as sources of hyperstable enzymes, for biomining of base and precious metals, and for evolutionary studies because of their use of eukaryotic-like subcellular mechanisms. Genetic methods are available for several species particularly Sulfolobus solfataricus. This organism has a considerable number of methods available for the construction of novel cell lines with unique functions. This chapter presents recent developments in the use of homologous recombination and linear DNA for the engineering of site-specific changes in the genome of S. solfataricus.

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