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WGS was performed to determine the genetic elements conferring antimicrobial resistance. Gene cloning, transcriptional analysis and targeted gene deletion, as well as protein purification and kinetic analysis, were performed to investigate the mechanism of resistance.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>APH(3\u2032)-IId consists of 266 amino acids and shares the highest identity (48.25%) with the previously known APH(3\u2032)-IIb. Expression of aph(3\u2032)-IId in Escherichia coli decreased susceptibility to kanamycin, neomycin, paromomycin and ribostamycin. The aph(3\u2032)-IId gene in ZJ499 was transcriptionally active under laboratory conditions and the relative abundance of this transcript was unaffected by treatment with the above four antibiotics. However, deletion of aph(3\u2032)-IId in ZJ499 results in decreased MICs of these drugs. The purified APH(3\u2032)-IId showed phosphotransferase activity against kanamycin, neomycin, paromomycin and ribostamycin, with catalytic efficiencies (kcat\/Km) ranging from \u223c105 to 107 M\u22121 s\u22121. Genetic environment and comparative genomic analyses suggested that aph(3\u2032)-IId is probably a ubiquitous gene in Brucella, with no mobile genetic elements detected in its surrounding region.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>APH(3\u2032)-IId is a novel chromosomal aminoglycoside phosphotransferase and plays an important role in the resistance of B. intermedia ZJ499 to kanamycin, neomycin, paromomycin and ribostamycin. To the best of our knowledge, APH(3\u2032)-IId represents the fourth characterized example of an APH(3\u2032)-II enzyme.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1093\/jac\/dkab272","type":"journal-article","created":{"date-parts":[[2021,7,13]],"date-time":"2021-07-13T19:09:41Z","timestamp":1626203381000},"page":"2787-2794","source":"Crossref","is-referenced-by-count":12,"title":["Identification and characteristics of a novel aminoglycoside phosphotransferase, APH(3\u2032)-IId, from an MDR clinical isolate of <i>Brucella intermedia<\/i>"],"prefix":"10.1093","volume":"76","author":[{"given":"Wei","family":"Lu","sequence":"first","affiliation":[{"name":"Key Laboratory of Medical Genetics of Zhejiang Province, Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China"},{"name":"Institute of Biomedical Informatics, School of Laboratory Medicine and Life Sciences, Wenzhou Medical 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