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Department of Microbiology, University of Georgia, Athens, Georgia 30602
* Corresponding author.
ABSTRACT
High-performance liquid chromatography is a promising alternative for determining the G+C content of bacterial deoxyribonucleic acid (DNA). The method which we evaluated involves enzymatic degradation of the DNA to nucleosides by P1 nuclease and bovine intestinal mucosa alkaline phosphatase, separation of the nucleosides by high-performance liquid chromatography, and calculation of the G+C content from the apparent ratios of deoxyguanosine and thymidine. Because the nucleosides are released from the DNA at different rates, incomplete degradation produces large errors in the apparent G+C content. For partially purified DNA, salts are a major source of interference in degradation. However, when the salts are carefully removed, the preparation and degradation of DNA contribute little error to the determination of G+C content. This method also requires careful selection of the chromatographic conditions to ensure separation of the major nucleosides from the nucleosides of modified bases and precise control of the flow rates. Both of these conditions are achievable with standard equipment and C18 reversed-phase columns. Then the method is precise, and the relative standard deviations of replicate measurements are close to 0.1%. It is also rapid, and a single measurement requires about 15 min. It requires small amounts of sample, and the G+C content can be determined from DNA isolated from a single bacterial colony. It is not affected by contamination with ribonucleic acid. Because this method yields a direct measurement, it may also be more accurate than indirect methods, such as the buoyant density and thermal denaturation methods. In addition, for highly purified DNA, the extent of modification can be determined.
Present address: Department of Pharmacognosy, Faculty of Pharmacy, University of Assiut, Assiut, Egypt. This article has been cited by other articles:
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Y.-X. Wang, Z.-G. Wang, J.-H. Liu, Y.-G. Chen, X.-X. Zhang, M.-L. Wen, L.-H. Xu, Q. Peng, and X.-L. Cui Sediminimonas qiaohouensis gen. nov., sp. nov., a member of the Roseobacter clade in the order Rhodobacterales Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1561 - 1567. [Abstract] [Full Text] [PDF] |
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C. Y. Hwang, G. D. Bae, W. Yih, and B. C. Cho Marivita cryptomonadis gen. nov., sp. nov. and Marivita litorea sp. nov., of the family Rhodobacteraceae, isolated from marine habitats Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1568 - 1575. [Abstract] [Full Text] [PDF] |
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R. H. Gregersen, C. Neubauer, H. Christensen, A. M. Bojesen, M. Hess, and M. Bisgaard Comparative studies on [Pasteurella] testudinis and [P.] testudinis-like bacteria and proposal of Chelonobacter oris gen. nov., sp. nov. as a new member of the family Pasteurellaceae Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1583 - 1588. [Abstract] [Full Text] [PDF] |
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R. Beaz Hidalgo, I. Cleenwerck, S. Balboa, S. Prado, P. De Vos, and J. L. Romalde Vibrio breoganii sp. nov., a non-motile, alginolytic, marine bacterium within the Vibrio halioticoli clade Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1589 - 1594. [Abstract] [Full Text] [PDF] |
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Y.-R. Cao, Y. Jiang, L.-H. Xu, and C.-L. Jiang Sphaerisporangium flaviroseum sp. nov. and Sphaerisporangium album sp. nov., isolated from forest soil in China Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1679 - 1684. [Abstract] [Full Text] [PDF] |
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S. Yoshizawa, M. Wada, K. Kita-Tsukamoto, E. Ikemoto, A. Yokota, and K. Kogure Vibrio azureus sp. nov., a luminous marine bacterium isolated from seawater Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1645 - 1649. [Abstract] [Full Text] [PDF] |
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Y.-G. Chen, Y.-Q. Zhang, J.-X. Shi, H.-D. Xiao, S.-K. Tang, Z.-X. Liu, K. Huang, X.-L. Cui, and W.-J. Li Jeotgalicoccus marinus sp. nov., a marine bacterium isolated from a sea urchin Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1625 - 1629. [Abstract] [Full Text] [PDF] |
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P. H. Summanen, P. A. Lawson, and S. M. Finegold Porphyromonas bennonis sp. nov., isolated from human clinical specimens Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1727 - 1732. [Abstract] [Full Text] [PDF] |
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H.-Y. Weon, R. Anandham, B.-Y. Kim, S.-B. Hong, Y.-A. Jeon, and S.-W. Kwon Dyella soli sp. nov. and Dyella terrae sp. nov., isolated from soil Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1685 - 1690. [Abstract] [Full Text] [PDF] |
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L.-P. Zhang, L.-M. Zhang, and X.-M. Zhang Streptosporangium canum sp. nov., isolated from soil Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1715 - 1719. [Abstract] [Full Text] [PDF] |
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N. S. Gordon, A. Valenzuela, S. M. Adams, P. W. Ramsey, J. L. Pollock, W. E. Holben, and J. E. Gannon Pedobacter nyackensis sp. nov., Pedobacter alluvionis sp. nov. and Pedobacter borealis sp. nov., isolated from Montana flood-plain sediment and forest soil Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1720 - 1726. [Abstract] [Full Text] [PDF] |
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Y.-G. Chen, Y.-Q. Zhang, H.-D. Xiao, Z.-X. Liu, L.-B. Yi, J.-X. Shi, X.-Y. Zhi, X.-L. Cui, and W.-J. Li Pontibacillus halophilus sp. nov., a moderately halophilic bacterium isolated from a sea urchin Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1635 - 1639. [Abstract] [Full Text] [PDF] |
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T. Clavel, C. Charrier, A. Braune, M. Wenning, M. Blaut, and D. Haller Isolation of bacteria from the ileal mucosa of TNFdeltaARE mice and description of Enterorhabdus mucosicola gen. nov., sp. nov. Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1805 - 1812. [Abstract] [Full Text] [PDF] |
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L. Zhang, Y. Wang, J. Dai, Y. Tang, Q. Yang, X. Luo, and C. Fang Bacillus korlensis sp. nov., a moderately halotolerant bacterium isolated from a sand soil sample in China Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1787 - 1792. [Abstract] [Full Text] [PDF] |
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I. Cleenwerck, M. De Wachter, A. Gonzalez, L. De Vuyst, and P. De Vos Differentiation of species of the family Acetobacteraceae by AFLP DNA fingerprinting: Gluconacetobacter kombuchae is a later heterotypic synonym of Gluconacetobacter hansenii Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1771 - 1786. [Abstract] [Full Text] [PDF] |
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S.-K. Tang, Y. Wang, K. Lou, P.-H. Mao, X. Jin, C.-L. Jiang, L.-H. Xu, and W.-J. Li Gracilibacillus saliphilus sp. nov., a moderately halophilic bacterium isolated from a salt lake Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1620 - 1624. [Abstract] [Full Text] [PDF] |
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F. Peng, M. Liu, L. Zhang, J. Dai, X. Luo, H. An, and C. Fang Planobacterium taklimakanense gen. nov., sp. nov., a member of the family Flavobacteriaceae that exhibits swimming motility, isolated from desert soil Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1672 - 1678. [Abstract] [Full Text] [PDF] |
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H.-Y. Weon, B.-Y. Kim, S.-B. Hong, Y.-A. Jeon, B.-S. Koo, S.-W. Kwon, and E. Stackebrandt Massilia niabensis sp. nov. and Massilia niastensis sp. nov., isolated from air samples Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1656 - 1660. [Abstract] [Full Text] [PDF] |
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J.-H. Kim, K.-Y. Kim, and C.-J. Cha Flavobacterium chungangense sp. nov., isolated from a freshwater lake Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1754 - 1758. [Abstract] [Full Text] [PDF] |
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P. Schumann, P. Kampfer, H.-J. Busse, L. I. Evtushenko, and for the Subcommittee on the Taxonomy of the Subord Proposed minimal standards for describing new genera and species of the suborder Micrococcineae Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1823 - 1849. [Abstract] [Full Text] [PDF] |
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E. V. Pikuta, R. B. Hoover, A. K. Bej, D. Marsic, W. B. Whitman, and P. Krader Spirochaeta dissipatitropha sp. nov., an alkaliphilic, obligately anaerobic bacterium, and emended description of the genus Spirochaeta Ehrenberg 1835 Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1798 - 1804. [Abstract] [Full Text] [PDF] |
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Y.-B. Ahn, J.-C. Chae, G. J. Zylstra, and M. M. Haggblom Degradation of Phenol via Phenylphosphate and Carboxylation to 4-Hydroxybenzoate by a Newly Isolated Strain of the Sulfate-Reducing Bacterium Desulfobacterium anilini Appl. Envir. Microbiol., July 1, 2009; 75(13): 4248 - 4253. [Abstract] [Full Text] [PDF] |
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H.-Y. Weon, S.-H. Yoo, Y.-J. Kim, J.-A Son, B.-Y. Kim, S.-W. Kwon, and B.-S. Koo Chitinophaga niabensis sp. nov. and Chitinophaga niastensis sp. nov., isolated from soil Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1267 - 1271. [Abstract] [Full Text] [PDF] |
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H.-Y. Weon, S.-H. Yoo, B.-Y. Kim, J.-A Son, Y.-J. Kim, and S.-W. Kwon Niabella ginsengisoli sp. nov., isolated from soil cultivated with Korean ginseng Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1282 - 1285. [Abstract] [Full Text] [PDF] |
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M. Liu, X. Luo, L. Zhang, J. Dai, Y. Wang, Y. Tang, J. Li, T. Sun, and C. Fang Pseudomonas xinjiangensis sp. nov., a moderately thermotolerant bacterium isolated from desert sand Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1286 - 1289. [Abstract] [Full Text] [PDF] |
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T.-W. Guan, S.-K. Tang, J.-Y. Wu, X.-Y. Zhi, L.-H. Xu, L.-L. Zhang, and W.-J. Li Haloglycomyces albus gen. nov., sp. nov., a halophilic, filamentous actinomycete of the family Glycomycetaceae Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1297 - 1301. [Abstract] [Full Text] [PDF] |
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K. Alain, N. Callac, M. Guegan, F. Lesongeur, P. Crassous, M.-A. Cambon-Bonavita, J. Querellou, and D. Prieur Nautilia abyssi sp. nov., a thermophilic, chemolithoautotrophic, sulfur-reducing bacterium isolated from an East Pacific Rise hydrothermal vent Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1310 - 1315. [Abstract] [Full Text] [PDF] |
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S.-K. Tang, Y. Wang, K. Lou, P.-H. Mao, L.-H. Xu, C.-L. Jiang, C.-J. Kim, and W.-J. Li Kocuria halotolerans sp. nov., an actinobacterium isolated from a saline soil in China Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1316 - 1320. [Abstract] [Full Text] [PDF] |
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Y.-G. Chen, X.-L. Cui, Q.-Y. Li, Y.-X. Wang, S.-K. Tang, Z.-X. Liu, M.-L. Wen, Q. Peng, and L.-H. Xu Saccharospirillum salsuginis sp. nov., a gammaproteobacterium from a subterranean brine Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1382 - 1386. [Abstract] [Full Text] [PDF] |
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L. Collado, I. Cleenwerck, S. Van Trappen, P. De Vos, and M. J. Figueras Arcobacter mytili sp. nov., an indoxyl acetate-hydrolysis-negative bacterium isolated from mussels Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1391 - 1396. [Abstract] [Full Text] [PDF] |
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D. W. Lee and S. D. Lee Dyella marensis sp. nov., isolated from cliff soil Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1397 - 1400. [Abstract] [Full Text] [PDF] |
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S. D. Lee Amycolatopsis ultiminotia sp. nov., isolated from rhizosphere soil, and emended description of the genus Amycolatopsis Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1401 - 1404. [Abstract] [Full Text] [PDF] |
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D. Wurdemann, B. J. Tindall, R. Pukall, H. Lunsdorf, C. Strompl, T. Namuth, H. Nahrstedt, M. Wos-Oxley, S. Ott, S. Schreiber, et al. Gordonibacter pamelaeae gen. nov., sp. nov., a new member of the Coriobacteriaceae isolated from a patient with Crohn's disease, and reclassification of Eggerthella hongkongensis Lau et al. 2006 as Paraeggerthella hongkongensis gen. nov., comb. nov. Int J Syst Evol Microbiol, June 1, 2009; 59(6): 1405 - 1415. [Abstract] [Full Text] [PDF] |
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