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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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K. Zhang, Y. Tang, L. Zhang, J. Dai, Y. Wang, X. Luo, M. Liu, G. Luo, and C. Fang Parasegetibacter luojiensis gen. nov., sp. nov., a member of the phylum Bacteroidetes isolated from a forest soil Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3058 - 3062. [Abstract] [Full Text] [PDF]
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C. Y. Hwang, G. I. Zhang, S.-H. Kang, H. J. Kim, and B. C. Cho Pseudomonas pelagia sp. nov., isolated from a culture of the Antarctic green alga Pyramimonas gelidicola Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3019 - 3024. [Abstract] [Full Text] [PDF]
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Y.-G. Chen, Y.-Q. Zhang, Y.-X. Wang, Z.-X. Liu, H.-P. Klenk, H.-D. Xiao, S.-K. Tang, X.-L. Cui, and W.-J. Li Bacillus neizhouensis sp. nov., a halophilic marine bacterium isolated from a sea anemone Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3035 - 3039. [Abstract] [Full Text] [PDF]
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J. Li, G.-Z. Zhao, S. Qin, H.-Y. Huang, W.-Y. Zhu, L.-H. Xu, and W.-J. Li Saccharopolyspora tripterygii sp. nov., an endophytic actinomycete isolated from the stem of Tripterygium hypoglaucum Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3040 - 3044. [Abstract] [Full Text] [PDF]
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Y.-S. Cui, S.-T. Lee, and W.-T. Im Nocardioides ginsengisoli sp. nov., isolated from soil of a ginseng field Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3045 - 3050. [Abstract] [Full Text] [PDF]
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S. H. Seo and S. D. Lee Actinocatenispora rupis sp. nov., isolated from cliff soil, and emended description of the genus Actinocatenispora Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3078 - 3082. [Abstract] [Full Text] [PDF]
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M. Halpern, T. Shaked, and P. Schumann Brachymonas chironomi sp. nov., isolated from a chironomid egg mass, and emended description of the genus Brachymonas Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3025 - 3029. [Abstract] [Full Text] [PDF]
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I. Vaz-Moreira, C. Faria, A. R. Lopes, L. Svensson, E. Falsen, E. R. B. Moore, A. C. S. Ferreira, O. C. Nunes, and C. M. Manaia Sphingobium vermicomposti sp. nov., isolated from vermicompost Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3145 - 3149. [Abstract] [Full Text] [PDF]
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Y. Wang, S.-K. Tang, K. Lou, J.-C. Lee, C. O. Jeon, L.-H. Xu, C.-J. Kim, and W.-J. Li Aidingimonas halophila gen. nov., sp. nov., a moderately halophilic bacterium isolated from a salt lake Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3088 - 3094. [Abstract] [Full Text] [PDF]
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N. Klouche, O. Basso, J.-F. Lascourreges, J.-L. Cayol, P. Thomas, G. Fauque, M.-L. Fardeau, and M. Magot Desulfocurvus vexinensis gen. nov., sp. nov., a sulfate-reducing bacterium isolated from a deep subsurface aquifer Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3100 - 3104. [Abstract] [Full Text] [PDF]
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J. Li, G.-Z. Zhao, S. Qin, W.-Y. Zhu, L.-H. Xu, and W.-J. Li Herbidospora osyris sp. nov., isolated from surface-sterilized tissue of Osyris wightiana Wall. ex Wight Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3123 - 3127. [Abstract] [Full Text] [PDF]
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K. Sucharita, Ch. Sasikala, S. C. Park, K. S. Baik, C. N. Seong, and Ch. V. Ramana Shewanella chilikensis sp. nov., a moderately alkaliphilic gammaproteobacterium isolated from a lagoon Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3111 - 3115. [Abstract] [Full Text] [PDF]
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J. Li, G.-Z. Zhao, H.-Y. Huang, S. Qin, W.-Y. Zhu, L.-H. Xu, and W.-J. Li Kineosporia mesophila sp. nov., isolated from surface-sterilized stems of Tripterygium wilfordii Int J Syst Evol Microbiol, December 1, 2009; 59(12): 3150 - 3154. [Abstract] [Full Text] [PDF]
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S.-C. Chang, M.-C. Wu, W.-M. Chen, Y.-H. Tsai, and T.-M. Lee Chitinilyticum litopenaei sp. nov., isolated from a freshwater shrimp pond, and emended description of the genus Chitinilyticum Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2651 - 2655. [Abstract] [Full Text] [PDF]
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I. D. Wagner, S. Ahmed, W. Zhao, C. L. Zhang, C. S. Romanek, M. Rohde, and J. Wiegel Caldanaerovirga acetigignens gen. nov., sp. nov., an anaerobic xylanolytic, alkalithermophilic bacterium isolated from Trego Hot Spring, Nevada, USA Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2685 - 2691. [Abstract] [Full Text] [PDF]
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Y.-S. Oh, H.-J. Lim, I.-T. Cha, W.-T. Im, J.-S. Yoo, U. G. Kang, S.-K. Rhee, and D.-H. Roh Roseovarius halotolerans sp. nov., isolated from deep seawater Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2718 - 2723. [Abstract] [Full Text] [PDF]
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Y.-G. Chen, Y.-X. Wang, Y.-Q. Zhang, S.-K. Tang, Z.-X. Liu, H.-D. Xiao, L.-H. Xu, X.-L. Cui, and W.-J. Li Nocardiopsis litoralis sp. nov., a halophilic marine actinomycete isolated from a sea anemone Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2708 - 2713. [Abstract] [Full Text] [PDF]
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M. K. Kim, S. Srinivasan, M.-J. Park, G. Sathiyaraj, Y.-J. Kim, and D.-C. Yang Nocardioides humi sp. nov., a {beta}-glucosidase-producing bacterium isolated from soil of a ginseng field Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2724 - 2728. [Abstract] [Full Text] [PDF]
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Y.-J. Lee, C. S. Romanek, and J. Wiegel Desulfosporosinus youngiae sp. nov., a spore-forming, sulfate-reducing bacterium isolated from a constructed wetland treating acid mine drainage Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2743 - 2746. [Abstract] [Full Text] [PDF]
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W. M. Moe, J. Yan, M. F. Nobre, M. S. da Costa, and F. A. Rainey Dehalogenimonas lykanthroporepellens gen. nov., sp. nov., a reductively dehalogenating bacterium isolated from chlorinated solvent-contaminated groundwater Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2692 - 2697. [Abstract] [Full Text] [PDF]
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S. Qin, G.-Z. Zhao, H.-P. Klenk, J. Li, W.-Y. Zhu, L.-H. Xu, and W.-J. Li Nonomuraea antimicrobica sp. nov., an endophytic actinomycete isolated from a leaf of Maytenus austroyunnanensis Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2747 - 2751. [Abstract] [Full Text] [PDF]
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A. B. Arun, W.-M. Chen, W.-A. Lai, J.-H. Chao, P. D. Rekha, F.-T. Shen, S. Singh, and C.-C. Young Muricauda lutaonensis sp. nov., a moderate thermophile isolated from a coastal hot spring Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2738 - 2742. [Abstract] [Full Text] [PDF]
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F. Wang, Y. Gai, M. Chen, and X. Xiao Arthrobacter psychrochitiniphilus sp. nov., a psychrotrophic bacterium isolated from Antarctica Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2759 - 2762. [Abstract] [Full Text] [PDF]
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C.-C. Young, S.-Y. Lin, A. B. Arun, F.-T. Shen, W.-M. Chen, P. D. Rekha, S. Langer, H.-J. Busse, Y.-H. Wu, and P. Kampfer Algoriphagus olei sp. nov., isolated from oil-contaminated soil Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2909 - 2915. [Abstract] [Full Text] [PDF]
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O. V. Golyshina, M. M. Yakimov, H. Lunsdorf, M. Ferrer, M. Nimtz, K. N. Timmis, V. Wray, B. J. Tindall, and P. N. Golyshin Acidiplasma aeolicum gen. nov., sp. nov., a euryarchaeon of the family Ferroplasmaceae isolated from a hydrothermal pool, and transfer of Ferroplasma cupricumulans to Acidiplasma cupricumulans comb. nov. Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2815 - 2823. [Abstract] [Full Text] [PDF]
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Y. Wang, L.-L. Cao, S.-K. Tang, K. Lou, P.-H. Mao, X. Jin, C.-L. Jiang, L.-H. Xu, and W.-J. Li Marinococcus luteus sp. nov., a halotolerant bacterium isolated from a salt lake, and emended description of the genus Marinococcus Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2875 - 2879. [Abstract] [Full Text] [PDF]
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X.-Q. Zhao, W.-J. Li, W.-C. Jiao, Y. Li, W.-J. Yuan, Y.-Q. Zhang, H.-P. Klenk, J.-W. Suh, and F.-W. Bai Streptomyces xinghaiensis sp. nov., isolated from marine sediment Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2870 - 2874. [Abstract] [Full Text] [PDF]
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D. W. Lee, J.-E. Lee, and S. D. Lee Chitinophaga rupis sp. nov., isolated from soil Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2830 - 2833. [Abstract] [Full Text] [PDF]
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Y.-G. Chen, Y.-Q. Zhang, H.-Y. Huang, H.-P. Klenk, S.-K. Tang, K. Huang, Q.-H. Chen, X.-L. Cui, and W.-J. Li Halomonas zhanjiangensis sp. nov., a halophilic bacterium isolated from a sea urchin Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2888 - 2893. [Abstract] [Full Text] [PDF]
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M. Madhaiyan, S. Poonguzhali, S.-W. Kwon, and T.-M. Sa Methylophilus rhizosphaerae sp. nov., a restricted facultative methylotroph isolated from rice rhizosphere soil Int J Syst Evol Microbiol, November 1, 2009; 59(11): 2904 - 2908. [Abstract] [Full Text] [PDF]
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G.-Z. Zhao, J. Li, S. Qin, Y.-Q. Zhang, W.-Y. Zhu, C.-L. Jiang, L.-H. Xu, and W.-J. Li Micrococcus yunnanensis sp. nov., a novel actinobacterium isolated from surface-sterilized Polyspora axillaris roots Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2383 - 2387. [Abstract] [Full Text] [PDF]
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J. H. Lim, S.-H. Baek, and S.-T. Lee Ferruginibacter alkalilentus gen. nov., sp. nov. and Ferruginibacter lapsinanis sp. nov., novel members of the family 'Chitinophagaceae' in the phylum Bacteroidetes, isolated from freshwater sediment Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2394 - 2399. [Abstract] [Full Text] [PDF]
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V. S. J. Schmidt, R. Mayr, M. Wenning, J. Glockner, H.-J. Busse, and S. Scherer Bavariicoccus seileri gen. nov., sp. nov., isolated from the surface and smear water of German red smear soft cheese Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2437 - 2443. [Abstract] [Full Text] [PDF]
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P. Kampfer, M. Vaneechoutte, N. Lodders, T. De Baere, V. Avesani, M. Janssens, H.-J. Busse, and G. Wauters Description of Chryseobacterium anthropi sp. nov. to accommodate clinical isolates biochemically similar to Kaistella koreensis and Chryseobacterium haifense, proposal to reclassify Kaistella koreensis as Chryseobacterium koreense comb. nov. and emended description of the genus Chryseobacterium Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2421 - 2428. [Abstract] [Full Text] [PDF]
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S. Qin, G.-Z. Zhao, J. Li, W.-Y. Zhu, L.-H. Xu, and W.-J. Li Actinomadura flavalba sp. nov., an endophytic actinomycete isolated from leaves of Maytenus austroyunnanensis Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2453 - 2457. [Abstract] [Full Text] [PDF]
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M. Cai, S.-K. Tang, Y.-G. Chen, Y. Li, Y.-Q. Zhang, and W.-J. Li Streptomonospora amylolytica sp. nov. and Streptomonospora flavalba sp. nov., two novel halophilic actinomycetes isolated from a salt lake Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2471 - 2475. [Abstract] [Full Text] [PDF]
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C.-Y. Wang, C.-C. Ng, W.-S. Tzeng, and Y.-T. Shyu Marinobacter szutsaonensis sp. nov., isolated from a solar saltern Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2605 - 2609. [Abstract] [Full Text] [PDF]
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Y.-X. Wang, J.-H. Liu, X.-X. Zhang, Y.-G. Chen, Z.-G. Wang, Y. Chen, Q.-Y. Li, Q. Peng, and X.-L. Cui Fodinicurvata sediminis gen. nov., sp. nov. and Fodinicurvata fenggangensis sp. nov., poly-{beta}-hydroxybutyrate-producing bacteria in the family Rhodospirillaceae Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2575 - 2581. [Abstract] [Full Text] [PDF]
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N. Vaisman and A. Oren Salisaeta longa gen. nov., sp. nov., a red, halophilic member of the Bacteroidetes Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2571 - 2574. [Abstract] [Full Text] [PDF]
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E. Ben-Dov, D. Z. Ben Yosef, V. Pavlov, and A. Kushmaro Corynebacterium maris sp. nov., a marine bacterium isolated from the mucus of the coral Fungia granulosa Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2458 - 2463. [Abstract] [Full Text] [PDF]
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Z. Ali, S. Cousin, A. Fruhling, E. Brambilla, P. Schumann, Y. Yang, and E. Stackebrandt Flavobacterium rivuli sp. nov., Flavobacterium subsaxonicum sp. nov., Flavobacterium swingsii sp. nov. and Flavobacterium reichenbachii sp. nov., isolated from a hard water rivulet Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2610 - 2617. [Abstract] [Full Text] [PDF]
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S.-Y. Sheu, T. F. Chiu, J.-H. Chou, D.-S. Sheu, A. B. Arun, C.-C. Young, C. A. Chen, J.-T. Wang, and W.-M. Chen Andreprevotia lacus sp. nov., isolated from a fish-culture pond Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2482 - 2485. [Abstract] [Full Text] [PDF]
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S.-Y. Sheu, T. F. Chiu, N.-T. Cho, J.-H. Chou, D.-S. Sheu, A. B. Arun, C.-C. Young, C. A. Chen, J.-T. Wang, and W.-M. Chen Flectobacillus roseus sp. nov., isolated from freshwater in Taiwan Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2546 - 2551. [Abstract] [Full Text] [PDF]
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A. B. Arun, C.-C. Young, W.-M. Chen, M.-H. Hung, W.-A. Lai, J.-H. Chou, P. D. Rekha, F.-T. Shen, and S. P. Su Belliella pelovolcani sp. nov., isolated from a mud-volcano in Taiwan Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2534 - 2537. [Abstract] [Full Text] [PDF]
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S. Qin, J. Li, Y.-Q. Zhang, W.-Y. Zhu, G.-Z. Zhao, L.-H. Xu, and W.-J. Li Plantactinospora mayteni gen. nov., sp. nov., a member of the family Micromonosporaceae Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2527 - 2533. [Abstract] [Full Text] [PDF]
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Y.-G. Chen, Y.-Q. Zhang, Z.-X. Liu, D.-C. Zhuang, H.-P. Klenk, S.-K. Tang, X.-L. Cui, and W.-J. Li Halobacillus salsuginis sp. nov., a moderately halophilic bacterium from a subterranean brine Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2505 - 2509. [Abstract] [Full Text] [PDF]
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J. Xiao, Y. Wang, Y. Luo, S.-J. Xie, J.-S. Ruan, and J. Xu Streptomyces avicenniae sp. nov., a novel actinomycete isolated from the rhizosphere of the mangrove plant Avicennia mariana Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2624 - 2628. [Abstract] [Full Text] [PDF]
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J.-H. Chou, Y.-J. Chou, A. B. Arun, C.-C. Young, C. A. Chen, J.-T. Wang, and W.-M. Chen Vogesella lacus sp. nov., isolated from a soft-shell turtle culture pond Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2629 - 2632. [Abstract] [Full Text] [PDF]
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C. Y. Hwang and B. C. Cho Spongiibacter tropicus sp. nov., isolated from a Synechococcus culture Int J Syst Evol Microbiol, September 1, 2009; 59(9): 2176 - 2179. [Abstract] [Full Text] [PDF]
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Y.-B. Ahn, L. J. Kerkhof, and M. M. Haggblom Desulfoluna spongiiphila sp. nov., a dehalogenating bacterium in the Desulfobacteraceae from the marine sponge Aplysina aerophoba Int J Syst Evol Microbiol, September 1, 2009; 59(9): 2133 - 2139. [Abstract] [Full Text] [PDF]
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Z.-G. Wang, Y.-X. Wang, J.-H. Liu, Y.-G. Chen, X.-X. Zhang, M.-L. Wen, L.-H. Xu, Q. Peng, and X.-L. Cui Fodinibacter luteus gen. nov., sp. nov., an actinobacterium isolated from a salt mine Int J Syst Evol Microbiol, September 1, 2009; 59(9): 2185 - 2190. [Abstract] [Full Text] [PDF]
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