{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T11:18:34Z","timestamp":1788520714129,"version":"build-2803163510"},"reference-count":55,"publisher":"Springer Science and Business Media LLC","issue":"1","content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Microbiol"],"published-print":{"date-parts":[[2008,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:sec>\n            <jats:title>Background<\/jats:title>\n            <jats:p>The sequenced genomes of the <jats:italic>Brucella<\/jats:italic> spp. have two urease operons, <jats:italic>ure<\/jats:italic>-1 and <jats:italic>ure<\/jats:italic>-2, but there is evidence that only one is responsible for encoding an active urease. The present work describes the purification and the enzymatic and phylogenomic characterization of urease from <jats:italic>Brucella suis<\/jats:italic> strain 1330. Additionally, the urease reactivity of sera from patients diagnosed with brucellosis was examined.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Results<\/jats:title>\n            <jats:p>Urease encoded by the <jats:italic>ure<\/jats:italic>-1 operon of <jats:italic>Brucella suis<\/jats:italic> strain 1330 was purified to homogeneity using ion exchange and hydrophobic interaction chromatographies. The urease was purified 51-fold with a recovery of 12% of the enzyme activity and 0.24% of the total protein. The enzyme had an isoelectric point of 5, and showed optimal activity at pH 7.0 and 28\u201335\u00b0C. The purified enzyme exhibited a Michaelis-Menten saturation kinetics with a <jats:italic>K<\/jats:italic>\n              <jats:sub>\n                <jats:italic>m<\/jats:italic>\n              <\/jats:sub>of 5.60 \u00b1 0.69 mM. Hydroxyurea and thiourea are competitive inhibitors of the enzyme with K<jats:sub>i<\/jats:sub> of 1.04 \u00b1 0.31 mM and 26.12 \u00b1 2.30 mM, respectively. Acetohydroxamic acid also inhibits the enzyme in a competitive way. The molecular weight estimated for the native enzyme was between 130\u2013135 kDa by gel filtration chromatography and 157 \u00b1 7 kDa using 5\u201310% polyacrylamide gradient non-denaturing gel. Only three subunits in SDS-PAGE were identified: two small subunits of 14,000 Da and 15,500 Da, and a major subunit of 66,000 Da. The amino terminal sequence of the purified large subunit corresponded to the predicted amino acid sequence encoded by <jats:italic>ureC1<\/jats:italic>. The UreC1 subunit was recognized by sera from patients with acute and chronic brucellosis. By phylogenetic and cluster structure analyses, <jats:italic>ureC1<\/jats:italic> was related to the <jats:italic>ureC<\/jats:italic> typically present in the <jats:italic>Rhizobiales<\/jats:italic>; in contrast, the <jats:italic>ureC2<\/jats:italic> encoded in the <jats:italic>ure<\/jats:italic>-2 operon is more related to distant species.<\/jats:p>\n          <\/jats:sec>\n          <jats:sec>\n            <jats:title>Conclusion<\/jats:title>\n            <jats:p>We have for the first time purified and characterized an active urease from <jats:italic>B. suis<\/jats:italic>. The enzyme was characterized at the kinetic, immunological and phylogenetic levels. Our results confirm that the active urease of <jats:italic>B. suis<\/jats:italic> is a product of <jats:italic>ure<\/jats:italic>-1 operon.<\/jats:p>\n          <\/jats:sec>","DOI":"10.1186\/1471-2180-8-121","type":"journal-article","created":{"date-parts":[[2008,7,19]],"date-time":"2008-07-19T06:13:44Z","timestamp":1216448024000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Enzymatic, immunological and phylogenetic characterization of Brucella suis urease"],"prefix":"10.1186","volume":"8","author":[{"given":"Araceli","family":"Contreras-Rodriguez","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose","family":"Quiroz-Limon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana M","family":"Martins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Humberto","family":"Peralta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric","family":"Avila-Calderon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nammalwar","family":"Sriranganathan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen M","family":"Boyle","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahide","family":"Lopez-Merino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2008,7,19]]},"reference":[{"issue":"3","key":"554_CR1","doi-asserted-by":"publisher","first-page":"621","DOI":"10.1111\/j.1365-2958.2004.04017.x","volume":"52","author":"RM Roop 2nd","year":"2004","unstructured":"Roop RM, Bellaire BH, Valderas MW, Cardelli JA: Adaptation of the Brucellae to their intracellular niche. Mol Microbiol. 2004, 52 (3): 621-630. 10.1111\/j.1365-2958.2004.04017.x.","journal-title":"Mol Microbiol"},{"issue":"7","key":"554_CR2","doi-asserted-by":"crossref","first-page":"3569","DOI":"10.1128\/jb.172.7.3569-3576.1990","volume":"172","author":"E Moreno","year":"1990","unstructured":"Moreno E, Stackebrandt E, Dorsch M, Wolters J, Busch M, Mayer H: Brucella abortus 16S rRNA and lipid A reveal a phylogenetic relationship with members of the alpha-2 subdivision of the class Proteobacteria. J Bacteriol. 1990, 172 (7): 3569-3576.","journal-title":"J Bacteriol"},{"key":"554_CR3","first-page":"867","volume-title":"Handbook of Metalloproteins","author":"RP Hausinger","year":"2001","unstructured":"Hausinger RP, Karplus PA: Urease. Handbook of Metalloproteins. Edited by: Wieghardt K, Huber R, Poulos TL, Messerschmidt A. 2001, West Sussex, John Wiley & Sons, Ltd, 867-879."},{"issue":"5","key":"554_CR4","doi-asserted-by":"publisher","first-page":"533","DOI":"10.1016\/S1286-4579(00)00312-9","volume":"2","author":"RA Burne","year":"2000","unstructured":"Burne RA, Chen YY: Bacterial ureases in infectious diseases. Microbes Infect. 2000, 2 (5): 533-542. 10.1016\/S1286-4579(00)00312-9.","journal-title":"Microbes Infect"},{"issue":"3","key":"554_CR5","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1128\/mr.59.3.451-480.1995","volume":"59","author":"HL Mobley","year":"1995","unstructured":"Mobley HL, Island MD, Hausinger RP: Molecular biology of microbial ureases. Microbiol Rev. 1995, 59 (3): 451-480.","journal-title":"Microbiol Rev"},{"issue":"5829","key":"554_CR6","doi-asserted-by":"publisher","first-page":"1307","DOI":"10.1126\/science.1139548","volume":"316","author":"E Giraud","year":"2007","unstructured":"Giraud E, Moulin L, Vallenet D, Barbe V, Cytryn E, Avarre JC, Jaubert M, Simon D, Cartieaux F, Prin Y, et al: Legumes symbioses: absence of Nod genes in photosynthetic bradyrhizobia. Science. 2007, 316 (5829): 1307-1312. 10.1126\/science.1139548.","journal-title":"Science"},{"key":"554_CR7","unstructured":"MBGD \u2013 Microbial Genome Database for Comparative Analysis. [http:\/\/mbgd.genome.ad.jp]"},{"issue":"16","key":"554_CR8","doi-asserted-by":"crossref","first-page":"9464","DOI":"10.1016\/S0021-9258(19)38872-6","volume":"265","author":"BE Dunn","year":"1990","unstructured":"Dunn BE, Campbell GP, Perez-Perez GI, Blaser MJ: Purification and characterization of urease from Helicobacter pylori. J Biol Chem. 1990, 265 (16): 9464-9469.","journal-title":"J Biol Chem"},{"issue":"4","key":"554_CR9","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1016\/0016-5085(88)90548-3","volume":"94","author":"CE Brady 3rd","year":"1988","unstructured":"Brady CE, Hadfield TL, Hyatt JR, Utts SJ: Acid secretion and serum gastrin levels in individuals with Campylobacter pylori. Gastroenterology. 1988, 94 (4): 923-927.","journal-title":"Gastroenterology"},{"issue":"10","key":"554_CR10","doi-asserted-by":"crossref","first-page":"3790","DOI":"10.1128\/iai.63.10.3790-3795.1995","volume":"63","author":"TF De Koning-Ward","year":"1995","unstructured":"De Koning-Ward TF, Robins-Browne RM: Contribution of urease to acid tolerance in Yersinia enterocolitica. Infect Immun. 1995, 63 (10): 3790-3795.","journal-title":"Infect Immun"},{"issue":"10","key":"554_CR11","doi-asserted-by":"crossref","first-page":"4507","DOI":"10.1128\/iai.61.10.4507-4509.1993","volume":"61","author":"P Probst","year":"1993","unstructured":"Probst P, Hermann E, Meyer zum Buschenfelde KH, Fleischer B: Identification of the Yersinia enterocolitica urease beta subunit as a target antigen for human synovial T lymphocytes in reactive arthritis. Infect Immun. 1993, 61 (10): 4507-4509.","journal-title":"Infect Immun"},{"key":"554_CR12","unstructured":"BRENDA \u2013 The Comprehensive Enzyme Information System. [http:\/\/www.brenda-enzymes.info\/]"},{"issue":"1","key":"554_CR13","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1016\/S0968-0004(01)02027-8","volume":"27","author":"I Schomburg","year":"2002","unstructured":"Schomburg I, Chang A, Hofmann O, Ebeling C, Ehrentreich F, Schomburg D: BRENDA: a resource for enzyme data and metabolic information. Trends Biochem Sci. 2002, 27 (1): 54-56. 10.1016\/S0968-0004(01)02027-8.","journal-title":"Trends Biochem Sci"},{"issue":"1","key":"554_CR14","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1093\/nar\/30.1.47","volume":"30","author":"I Schomburg","year":"2002","unstructured":"Schomburg I, Chang A, Schomburg D: BRENDA, enzyme data and metabolic information. Nucleic Acids Res. 2002, 30 (1): 47-49. 10.1093\/nar\/30.1.47.","journal-title":"Nucleic Acids Res"},{"key":"554_CR15","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1016\/B978-044452085-2\/50004-8","volume-title":"Systems biology: philosophical foundations","author":"HV Westerhoff","year":"2007","unstructured":"Westerhoff HV, Kell DB: The methodologies of systems biology. Systems biology: philosophical foundations. Edited by: Boogerd FC, Hofmeyer JH, Westerhoff HV. 2007, Amsterdam, Elsevier, 23-70."},{"issue":"19","key":"554_CR16","doi-asserted-by":"crossref","first-page":"5644","DOI":"10.1128\/jb.177.19.5644-5652.1995","volume":"177","author":"DL Clemens","year":"1995","unstructured":"Clemens DL, Lee BY, Horwitz MA: Purification, characterization, and genetic analysis of Mycobacterium tuberculosis urease, a potentially critical determinant of host-pathogen interaction. J Bacteriol. 1995, 177 (19): 5644-5652.","journal-title":"J Bacteriol"},{"key":"554_CR17","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1007\/BF01570869","volume":"27","author":"M del Mar Dobao","year":"1993","unstructured":"del Mar Dobao M, Castrillo F, Pineda M: Characterization of urease from the phototropic bacterium Rhodobacter capsulatus E1F1. Curr Microbiol. 1993, 27: 119-123. 10.1007\/BF01570869.","journal-title":"Curr Microbiol"},{"key":"554_CR18","doi-asserted-by":"publisher","first-page":"1495","DOI":"10.1271\/bbb1961.48.1495","volume":"48","author":"HTS Nakano","year":"1984","unstructured":"Nakano HTS, Watanabe Y: Purification and properties of urease from Brevibacterium ammoniagenes. Agric Biol Chem. 1984, 48: 1495-1502.","journal-title":"Agric Biol Chem"},{"key":"554_CR19","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1271\/bbb1961.54.381","volume":"54","author":"S Kakimoto","year":"1990","unstructured":"Kakimoto S, Miyashita H, Sumino Y, Akiyama S: Properties of acid ureases from Lactobacilus and Streptococcus strains. Agric Biol Chem. 1990, 54: 381-386.","journal-title":"Agric Biol Chem"},{"issue":"12","key":"554_CR20","doi-asserted-by":"crossref","first-page":"2302","DOI":"10.1128\/jcm.25.12.2302-2305.1987","volume":"25","author":"HL Mobley","year":"1987","unstructured":"Mobley HL, Jones BD, Penner JL: Urease activity of Proteus penneri. J Clin Microbiol. 1987, 25 (12): 2302-2305.","journal-title":"J Clin Microbiol"},{"issue":"2","key":"554_CR21","doi-asserted-by":"publisher","first-page":"774","DOI":"10.1128\/IAI.01244-06","volume":"75","author":"FJ Sangari","year":"2007","unstructured":"Sangari FJ, Seoane A, Rodriguez MC, Aguero J, Garcia Lobo JM: Characterization of the urease operon of Brucella abortus and assessment of its role in virulence of the bacterium. Infect Immun. 2007, 75 (2): 774-780. 10.1128\/IAI.01244-06.","journal-title":"Infect Immun"},{"issue":"6","key":"554_CR22","doi-asserted-by":"publisher","first-page":"487","DOI":"10.1139\/m86-089","volume":"32","author":"H Eng","year":"1986","unstructured":"Eng H, Robertson JA, Stemke GW: Properties of urease from Ureaplasma urealyticum: kinetics, molecular weight, and demonstration of multiple enzyme isoelectric point forms. Can J Microbiol. 1986, 32 (6): 487-493.","journal-title":"Can J Microbiol"},{"issue":"5","key":"554_CR23","first-page":"393","volume":"161","author":"UK Schafer","year":"1994","unstructured":"Schafer UK, Kaltwasser H: Urease from Staphylococcus saprophyticus: purification, characterization and comparison to Staphylococcus xylosus urease. Arch Microbiol. 1994, 161 (5): 393-399.","journal-title":"Arch Microbiol"},{"issue":"29","key":"554_CR24","doi-asserted-by":"publisher","first-page":"8575","DOI":"10.1021\/bi000613o","volume":"39","author":"MA Pearson","year":"2000","unstructured":"Pearson MA, Park IS, Schaller RA, Michel LO, Karplus PA, Hausinger RP: Kinetic and structural characterization of urease active site variants. Biochemistry. 2000, 39 (29): 8575-8584. 10.1021\/bi000613o.","journal-title":"Biochemistry"},{"issue":"9","key":"554_CR25","doi-asserted-by":"crossref","first-page":"2198","DOI":"10.1128\/iai.55.9.2198-2203.1987","volume":"55","author":"BD Jones","year":"1987","unstructured":"Jones BD, Mobley HL: Genetic and biochemical diversity of ureases of Proteus, Providencia, and Morganella species isolated from urinary tract infection. Infect Immun. 1987, 55 (9): 2198-2203.","journal-title":"Infect Immun"},{"issue":"1","key":"554_CR26","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1128\/mr.53.1.85-108.1989","volume":"53","author":"HL Mobley","year":"1989","unstructured":"Mobley HL, Hausinger RP: Microbial ureases: significance, regulation, and molecular characterization. Microbiol Rev. 1989, 53 (1): 85-108.","journal-title":"Microbiol Rev"},{"issue":"27","key":"554_CR27","doi-asserted-by":"crossref","first-page":"15835","DOI":"10.1016\/S0021-9258(18)71553-6","volume":"264","author":"MJ Todd","year":"1989","unstructured":"Todd MJ, Hausinger RP: Competitive inhibitors of Klebsiella aerogenes urease. Mechanisms of interaction with the nickel active site. J Biol Chem. 1989, 264 (27): 15835-15842.","journal-title":"J Biol Chem"},{"issue":"10","key":"554_CR28","doi-asserted-by":"publisher","first-page":"6264","DOI":"10.1128\/IAI.69.10.6264-6270.2001","volume":"69","author":"A Al-Mariri","year":"2001","unstructured":"Al-Mariri A, Tibor A, Mertens P, De Bolle X, Michel P, Godfroid J, Walravens K, Letesson JJ: Induction of immune response in BALB\/c mice with a DNA vaccine encoding bacterioferritin or P39 of Brucella spp. Infect Immun. 2001, 69 (10): 6264-6270. 10.1128\/IAI.69.10.6264-6270.2001.","journal-title":"Infect Immun"},{"issue":"1","key":"554_CR29","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1128\/jcm.34.1.165-169.1996","volume":"34","author":"OL Rossetti","year":"1996","unstructured":"Rossetti OL, Arese AI, Boschiroli ML, Cravero SL: Cloning of Brucella abortus gene and characterization of expressed 26-kilodalton periplasmic protein: potential use for diagnosis. J Clin Microbiol. 1996, 34 (1): 165-169.","journal-title":"J Clin Microbiol"},{"issue":"5","key":"554_CR30","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1128\/cdli.1.5.506-510.1994","volume":"1","author":"LB Tabatabai","year":"1994","unstructured":"Tabatabai LB, Hennager SG: Cattle serologically positive for Brucella abortus have antibodies to B. abortus Cu-Zn superoxide dismutase. Clin Diagn Lab Immunol. 1994, 1 (5): 506-510.","journal-title":"Clin Diagn Lab Immunol"},{"key":"554_CR31","doi-asserted-by":"publisher","first-page":"57","DOI":"10.1186\/1471-2180-7-57","volume":"7","author":"AB Bandara","year":"2007","unstructured":"Bandara AB, Contreras A, Contreras-Rodriguez A, Martins AM, Dobrean V, Poff-Reichow S, Rajasekaran P, Sriranganathan N, Schurig GG, Boyle SM: Brucella suis urease encoded by ure1 but not ure2 is necessary for intestinal infection of BALB\/c mice. BMC Microbiol. 2007, 7: 57-10.1186\/1471-2180-7-57.","journal-title":"BMC Microbiol"},{"issue":"9","key":"554_CR32","first-page":"1964","volume":"26","author":"H Appel","year":"1999","unstructured":"Appel H, Mertz A, Distler A, Sieper J, Braun J: The 19 kDa protein of Yersinia enterocolitica O:3 is recognized on the cellular and humoral level by patients with Yersinia induced reactive arthritis. J Rheumatol. 1999, 26 (9): 1964-1971.","journal-title":"J Rheumatol"},{"issue":"5","key":"554_CR33","first-page":"207","volume":"26","author":"JJ Dubost","year":"1997","unstructured":"Dubost JJ, Constantin A, Soubrier M, Ristori JM, Cantagrel A, Bussiere JL: [Does reactive arthritis caused by Brucella exist? Apropos of 4 cases]. Presse Med. 1997, 26 (5): 207-210.","journal-title":"Presse Med"},{"issue":"1","key":"554_CR34","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1034\/j.1600-0420.2001.079001076.x","volume":"79","author":"K Gungor","year":"2001","unstructured":"Gungor K, Bekir NA, Namiduru M: Recurrent episcleritis associated with brucellosis. Acta Ophthalmol Scand. 2001, 79 (1): 76-78. 10.1034\/j.1600-0420.2001.079001076.x.","journal-title":"Acta Ophthalmol Scand"},{"issue":"2","key":"554_CR35","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1016\/S1521-6942(02)00104-3","volume":"17","author":"JS Hill-Gaston","year":"2003","unstructured":"Hill-Gaston JS, Lillicrap MS: Arthritis associated with enteric infection. Best Pract Res Clin Rheumatol. 2003, 17 (2): 219-239. 10.1016\/S1521-6942(02)00104-3.","journal-title":"Best Pract Res Clin Rheumatol"},{"issue":"3","key":"554_CR36","doi-asserted-by":"publisher","first-page":"892","DOI":"10.1002\/art.20948","volume":"52","author":"P Atagunduz","year":"2005","unstructured":"Atagunduz P, Appel H, Kuon W, Wu P, Thiel A, Kloetzel PM, Sieper J: HLA-B27-restricted CD8+ T cell response to cartilage-derived self peptides in ankylosing spondylitis. Arthritis Rheum. 2005, 52 (3): 892-901. 10.1002\/art.20948.","journal-title":"Arthritis Rheum"},{"issue":"4","key":"554_CR37","doi-asserted-by":"crossref","first-page":"1315","DOI":"10.1128\/MMBR.62.4.1315-1352.1998","volume":"62","author":"GR Cornelis","year":"1998","unstructured":"Cornelis GR, Boland A, Boyd AP, Geuijen C, Iriarte M, Neyt C, Sory MP, Stainier I: The virulence plasmid of Yersinia, an antihost genome. Microbiol Mol Biol Rev. 1998, 62 (4): 1315-1352.","journal-title":"Microbiol Mol Biol Rev"},{"issue":"9","key":"554_CR38","doi-asserted-by":"publisher","first-page":"5238","DOI":"10.1128\/IAI.71.9.5238-5244.2003","volume":"71","author":"A Contreras-Rodriguez","year":"2003","unstructured":"Contreras-Rodriguez A, Ramirez-Zavala B, Contreras A, Schurig GG, Sriranganathan N, Lopez-Merino A: Purification and characterization of an immunogenic aminopeptidase of Brucella melitensis. Infect Immun. 2003, 71 (9): 5238-5244. 10.1128\/IAI.71.9.5238-5244.2003.","journal-title":"Infect Immun"},{"issue":"5","key":"554_CR39","first-page":"327","volume":"16","author":"JM Hamilton-Miller","year":"1979","unstructured":"Hamilton-Miller JM, Gargan RA: Rapid screening for urease inhibitors. Invest Urol. 1979, 16 (5): 327-328.","journal-title":"Invest Urol"},{"issue":"1","key":"554_CR40","doi-asserted-by":"publisher","first-page":"132","DOI":"10.1016\/0003-2697(66)90088-1","volume":"16","author":"H Kaltwasser","year":"1966","unstructured":"Kaltwasser H, Schlegel HG: NADH-Dependent coupled enzyme assay for urease and other ammonia-producing systems. Anal Biochem. 1966, 16 (1): 132-138. 10.1016\/0003-2697(66)90088-1.","journal-title":"Anal Biochem"},{"issue":"2","key":"554_CR41","doi-asserted-by":"publisher","first-page":"321","DOI":"10.1006\/abio.1994.1591","volume":"223","author":"CA Cordeiro","year":"1994","unstructured":"Cordeiro CA, Freire AP: Protein determination in permeabilized yeast cells using the Coomassie brilliant blue dye binding assay. Anal Biochem. 1994, 223 (2): 321-323. 10.1006\/abio.1994.1591.","journal-title":"Anal Biochem"},{"issue":"5259","key":"554_CR42","doi-asserted-by":"publisher","first-page":"680","DOI":"10.1038\/227680a0","volume":"227","author":"UK Laemmli","year":"1970","unstructured":"Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970, 227 (5259): 680-685. 10.1038\/227680a0.","journal-title":"Nature"},{"issue":"21","key":"554_CR43","doi-asserted-by":"crossref","first-page":"10035","DOI":"10.1016\/S0021-9258(18)61070-1","volume":"262","author":"P Matsudaira","year":"1987","unstructured":"Matsudaira P: Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987, 262 (21): 10035-10038.","journal-title":"J Biol Chem"},{"key":"554_CR44","unstructured":"NCBI \u2013 National Center for Biotechnology Information. [http:\/\/www.ncbi.nlm.nih.gov\/]"},{"issue":"17","key":"554_CR45","doi-asserted-by":"publisher","first-page":"3389","DOI":"10.1093\/nar\/25.17.3389","volume":"25","author":"SF Altschul","year":"1997","unstructured":"Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ: Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997, 25 (17): 3389-3402. 10.1093\/nar\/25.17.3389.","journal-title":"Nucleic Acids Res"},{"key":"554_CR46","volume-title":"HyperFit \u2013 Hyperbolic Regression Analysis of Enzyme Kinetic Data. (version 1.1)","author":"JS Easterby","year":"1993","unstructured":"Easterby JS: HyperFit \u2013 Hyperbolic Regression Analysis of Enzyme Kinetic Data. (version 1.1). 1993"},{"key":"554_CR47","volume-title":"Fundamentals of Enzyme Kinetics","author":"A Cornish-Bowden","year":"1995","unstructured":"Cornish-Bowden A: Fundamentals of Enzyme Kinetics. 1995, London, Portland Press"},{"issue":"9","key":"554_CR48","doi-asserted-by":"publisher","first-page":"4350","DOI":"10.1073\/pnas.76.9.4350","volume":"76","author":"H Towbin","year":"1979","unstructured":"Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Nat Acad Sci USA. 1979, 76 (9): 4350-4354. 10.1073\/pnas.76.9.4350.","journal-title":"Proc Nat Acad Sci USA"},{"issue":"22","key":"554_CR49","doi-asserted-by":"publisher","first-page":"4673","DOI":"10.1093\/nar\/22.22.4673","volume":"22","author":"JD Thompson","year":"1994","unstructured":"Thompson JD, Higgins DG, Gibson TJ: CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994, 22 (22): 4673-4680. 10.1093\/nar\/22.22.4673.","journal-title":"Nucleic Acids Res"},{"issue":"4","key":"554_CR50","first-page":"406","volume":"4","author":"N Saitou","year":"1987","unstructured":"Saitou N, Nei M: The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987, 4 (4): 406-425.","journal-title":"Mol Biol Evol"},{"key":"554_CR51","first-page":"164","volume":"5","author":"J Felsenstein","year":"1989","unstructured":"Felsenstein J: PHYLIP \u2013 Phylogeny Inference Package (version 3.2). Cladistics. 1989, 5: 164-166.","journal-title":"Cladistics"},{"issue":"5","key":"554_CR52","doi-asserted-by":"publisher","first-page":"696","DOI":"10.1080\/10635150390235520","volume":"52","author":"S Guindon","year":"2003","unstructured":"Guindon S, Gascuel O: A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol. 2003, 52 (5): 696-704. 10.1080\/10635150390235520.","journal-title":"Syst Biol"},{"issue":"4","key":"554_CR53","first-page":"357","volume":"12","author":"RD Page","year":"1996","unstructured":"Page RD: TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci. 1996, 12 (4): 357-358.","journal-title":"Comput Appl Biosci"},{"key":"554_CR54","unstructured":"RhizoBase \u2013 The Genome Database for Rhizobia. [http:\/\/bacteria.kazusa.or.jp\/rhizobase\/]"},{"key":"554_CR55","unstructured":"KEGG \u2013 Kyoto Encyclopedia of Genes and Genomes. [http:\/\/www.genome.jp\/kegg\/]"}],"container-title":["BMC Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/1471-2180-8-121.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,9,1]],"date-time":"2021-09-01T10:09:30Z","timestamp":1630490970000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcmicrobiol.biomedcentral.com\/articles\/10.1186\/1471-2180-8-121"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2008,7,19]]},"references-count":55,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2008,12]]}},"alternative-id":["554"],"URL":"https:\/\/doi.org\/10.1186\/1471-2180-8-121","relation":{},"ISSN":["1471-2180"],"issn-type":[{"value":"1471-2180","type":"electronic"}],"subject":[],"published":{"date-parts":[[2008,7,19]]},"assertion":[{"value":"13 November 2007","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 July 2008","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 July 2008","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}}],"article-number":"121"}}