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Antimicrob Agents Chemother. 1986 August; 30(2): 315–320.
PMCID: PMC180541
Molecular cloning of amikacin resistance determinants from a Klebsiella pneumoniae plasmid.
M E Tolmasky, M Roberts, M Woloj, and J H Crosa
Abstract
A multiresistant Klebsiella pneumoniae strain, JHCK1, harbored several plasmids. One of these, plasmid pJHCMW1, carried determinants for resistance to amikacin in addition to kanamycin, tobramycin, and ampicillin. The amikacin resistance determinant(s) was cloned and studied by restriction mapping, insertion, and deletion analysis. The amikacin resistance gene(s) was localized in a 1.5-kilobase DNA fragment. This pJHCMW1 DNA region was responsible for not only the resistance to amikacin but also the resistance to kanamycin and tobramycin. The cloned DNA fragment specified both an acetyltransferase activity and a low level of phosphotransferase activity. The two activities were absent from mutants that did not confer resistance to amikacin, kanamycin, and tobramycin.
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Selected References
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  • Bachmann BJ. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. [PubMed]
  • Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966 Apr;45(4):493–496. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PubMed]
  • Bolivar F. Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA molecules. Gene. 1978 Oct;4(2):121–136. [PubMed]
  • Bongaerts GP, Kaptijn GM. Aminoglycoside phosphotransferase-II-mediated amikacin resistance in Escherichia coli. Antimicrob Agents Chemother. 1981 Sep;20(3):344–350. [PubMed]
  • Boyer HW, Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. [PubMed]
  • Chang AC, Cohen SN. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. [PubMed]
  • Cohen SN, Chang AC, Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. [PubMed]
  • Coombe RG, George AM. New plasmid-mediated aminoglycoside adenylyltransferase of broad substrate range that adenylylates amikacin. Antimicrob Agents Chemother. 1981 Jul;20(1):75–80. [PubMed]
  • Crosa JH, Actis LA, Mitoma Y, Perez-Casal J, Tolmasky ME, Valvano MA. Plasmid-mediated iron sequestering systems in pathogenic strains of Vibrio anguillarum and Escherichia coli. Basic Life Sci. 1985;30:759–774. [PubMed]
  • Crosa JH, Olarte J, Mata LJ, Luttropp LK, Peñaranda ME. Characterization of an R-plasmid associated with ampicillin resistance in Shigella dysenteriae type 1 isolated from epidemics. Antimicrob Agents Chemother. 1977 Mar;11(3):553–558. [PubMed]
  • Crosa JH, Schiewe MH, Falkow S. Evidence for plasmid contribution to the virulence of fish pathogen Vibrio anguillarum. Infect Immun. 1977 Nov;18(2):509–513. [PubMed]
  • Davies J, Smith DI. Plasmid-determined resistance to antimicrobial agents. Annu Rev Microbiol. 1978;32:469–518. [PubMed]
  • Guiney DG, Helinski DR. The DNA-protein relaxation complex of the plasmid RK2: location of the site-specific nick in the region of the proposed origin of transfer. Mol Gen Genet. 1979 Oct 3;176(2):183–189. [PubMed]
  • Haas MJ, Dowding JE. Aminoglycoside-modifying enzymes. Methods Enzymol. 1975;43:611–628. [PubMed]
  • Jacoby GA. Properties of an R plasmid in Pseudomonas aeruginosa producing amikacin (BB-K8), butirosin, kanamycin, tobramycin, and sisomicin resistance. Antimicrob Agents Chemother. 1974 Dec;6(6):807–810. [PubMed]
  • John JF, Jr, McNeill WF, Price KE, Kresel PA. Evidence for a chromosomal site specifying amikacin resistance in multiresistant Serratia marcescens. Antimicrob Agents Chemother. 1982 Apr;21(4):587–591. [PubMed]
  • Le Goffic F, Moreau N, Masson M. Are some aminoglycoside antibiotics inactivating enzymes polyfunctional? Ann Microbiol (Paris). 1977 Nov–Dec;128B(4):465–469. [PubMed]
  • Le Goffic F, Moreau N, Soussy CJ, Duval J. How can DKB and tobramycin be inactivated on two different sites by the same enzyme? Ann Microbiol (Paris). 1977 Nov–Dec;128B(4):459–463. [PubMed]
  • Levine JF, Maslow MJ, Leibowitz RE, Pollock AA, Hanna BA, Schaefler S, Simberkoff MS, Rahal JJ., Jr Amikacin-resistant gram-negative bacilli: correlation of occurrence with amikacin use. J Infect Dis. 1985 Feb;151(2):295–300. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Macrina FL, Kopecko DJ, Jones KR, Ayers DJ, McCowen SM. A multiple plasmid-containing Escherichia coli strain: convenient source of size reference plasmid molecules. Plasmid. 1978 Jun;1(3):417–420. [PubMed]
  • Meyer JF, Nies BA, Wiedemann B. Amikacin resistance mediated by multiresistance transposon Tn2424. J Bacteriol. 1983 Aug;155(2):755–760. [PubMed]
  • Murray BE, Moellering RC., Jr In-vivo acquisition of two different types of aminoglycoside resistance by a single strain of Klebsiella pneumoniae causing severe infection. Ann Intern Med. 1982 Feb;96(2):176–180. [PubMed]
  • Perlin MH, Lerner SA. Amikacin resistance associated with a plasmid-borne aminoglycoside phosphotransferase in Escherichia coli. Antimicrob Agents Chemother. 1979 Nov;16(5):598–604. [PubMed]
  • Radika K, Northrop DB. Correlation of antibiotic resistance with Vmax/Km ratio of enzymatic modification of aminoglycosides by kanamycin acetyltransferase. Antimicrob Agents Chemother. 1984 Apr;25(4):479–482. [PubMed]
  • Rubens CE, McNeill WF, Farrar WE., Jr Evolution of multiple-antibiotic-resistance plasmids mediated by transposable plasmid deoxyribonucleic acid sequences. J Bacteriol. 1979 Nov;140(2):713–719. [PubMed]
  • Tenover FC, Gootz TD, Gordon KP, Tompkins LS, Young SA, Plorde JJ. Development of a DNA probe for the structural gene of the 2"-O-adenyltransferase aminoglycoside-modifying enzyme. J Infect Dis. 1984 Nov;150(5):678–687. [PubMed]
  • Woloj M, Tolmasky ME, Roberts MC, Crosa JH. Plasmid-encoded amikacin resistance in multiresistant strains of Klebsiella pneumoniae isolated from neonates with meningitis. Antimicrob Agents Chemother. 1986 Feb;29(2):315–319. [PubMed]
  • Young SA, Tenover FC, Gootz TD, Gordon KP, Plorde JJ. Development of two DNA probes for differentiating the structural genes of subclasses I and II of the aminoglycoside-modifying enzyme 3'-aminoglycoside phosphotransferase. Antimicrob Agents Chemother. 1985 May;27(5):739–744. [PubMed]