PMID: 11607093Aug 1, 1990Paper

Method for isolation of auxotrophs in the methanogenic archaebacteria: role of the acetyl-CoA pathway of autotrophic CO2 fixation in Methanococcus maripaludis

Proceedings of the National Academy of Sciences of the United States of America
J Ladapo, William B Whitman

Abstract

A procedure was developed for the enrichment of auxotrophs in the antibiotic-insensitive archaebacterium Methanococcus. After mutagenesis with ethyl methanesulfonate, growing cells were selectively killed upon exposure to the base analogs 6-azauracil and 8-azahypoxanthine for 48 hr. Using this method, eight independent acetate autotrophs of Methanococcus maripaludis were isolated. Six of the auxotrophs had an absolute growth requirement for acetate and contained 1-16% of the wild-type levels of CO dehydrogenase. Three of these six also contained 14-29% of the wild-type levels of pyruvate oxidoreductase and 12-30% of the wild-type levels of pyruvate synthase. Two spontaneous revertants of these latter auxotrophs regained the ability to grow normally in the absence of acetate and wild-type levels of CO dehydrogenase, acetyl-CoA synthase, pyruvate oxidoreductase, and pyruvate synthase. Likewise, a spontaneous revertant of an auxotroph with reduced levels of CO dehydrogenase and wild-type levels of pyruvate oxidoreductase regained the ability to grow normally in the absence of acetate and wild-type levels of CO dehydrogenase and acetyl-CoA synthase. Two additional auxotrophs grew poorly in the absence of acetate but contained wild-...Continue Reading

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Citations

Jun 8, 2007·Archives of Microbiology·Michael RotherWilliam M Metcalf
Jun 26, 2008·Archives of Microbiology·Ellen Oelgeschläger, Michael Rother
Jul 8, 2011·Archives of Microbiology·Michael KliefothMichael Rother
Jan 5, 2005·Nature Reviews. Genetics·Thorsten Allers, Moshe Mevarech
Jun 11, 2010·Proceedings of the National Academy of Sciences of the United States of America·Kyle C CostaJohn A Leigh
Apr 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·D R Abbanat, J G Ferry
Jul 16, 2008·Proceedings of the National Academy of Sciences of the United States of America·Weimin GongMichael K Chan
Mar 18, 1997·Proceedings of the National Academy of Sciences of the United States of America·W W MetcalfR S Wolfe
Nov 20, 2004·Proceedings of the National Academy of Sciences of the United States of America·Michael Rother, William W Metcalf
Apr 16, 2014·Proceedings of the National Academy of Sciences of the United States of America·Wei-Qin ZhuangLisa Alvarez-Cohen
Feb 1, 2011·Metabolic Engineering·Nanette R Boyle, John A Morgan
Aug 11, 2015·Biochimica Et Biophysica Acta·Jessica Hadj-SaïdSébastien Dementin
Jul 18, 2008·Environmental Microbiology·Elizabeth PierceStephen W Ragsdale
Nov 1, 2005·Current Opinion in Microbiology·Michael Rother, William W Metcalf
Jan 1, 1991·Critical Reviews in Biochemistry and Molecular Biology·S W Ragsdale
Jun 12, 2016·Microbial Cell Factories·Nishu GoyalIftekhar A Karimi
May 14, 2003·Archives of Microbiology·Winston C LinWilliam B Whitman
Jan 20, 2018·Archaea : an International Microbiological Journal·Feng LongWilliam B Whitman
Jan 20, 2019·Applied and Environmental Microbiology·Sara KleindienstFrank E Löffler
Jul 20, 1999·Molecular Microbiology·D L Tumbula, W B Whitman
Dec 12, 2018·Frontiers in Microbiology·Nicole Adam, Mirjam Perner
Jun 12, 2003·Chemical Reviews·Stephen W Ragsdale
Sep 1, 1992·Journal of Bacteriology·J G Ferry
Sep 20, 2008·Biochimica Et Biophysica Acta·Stephen W Ragsdale, Elizabeth Pierce

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