Summary
Molecular cloning of DNA fragments between 1.5 and 8kb from BamHI, EcoRI, HindIII, SalI, or Sau3A digests permitted the isolation of structural genes coding for TEM-1, ROB-1, OXA-1, OXA-3, OXA-4, OXA-5, PSE-1, PSE-2, PSE-3, PSE-4, CARB-3, CARB-4, AER-1, and LCR-1 β-lactamases. Ampicillin-resistant clones were selected and it was confirmed that they contained the respective β-lactamase genes by isoelectric focusing. Detailed physical maps of 14 different recombinant plasmids were constructed using 8 restriction endonucleases. Plasmid deletions and lacZ fusions were used to localize the β-lactamase structural genes. DNA probes were constructed for the TEM01, ROB-1, OXA-1, and PSE-1 genes. Under conditions of high stringency, hybridization was observed between the genes for TEM-1 and TEM-2 or TLE-1, OXA-1 and OXA-4, and PSE-1 and PSE-4 or CARB-3, while the ROB-1 gene probe showed no cross-hybridization. Such bla gene probes should facilitate studies of β-lactamase molecular epidemiology.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ambler RP (1980) The structure of β-lactamases. Philos Trans R Soc Lond B 289:321–331
Ambler RP, Scott GK (1978) Partial amino acid sequence of penicllinase coded by Escherichia coli plasmid R6K. Proc Natl Acad Sci USA 75:3732–3736
Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Bolivar F, Backman K (1979) Plasmids of Escherichia coli as cloning vectors. Methods Enzymol 68:245–267
Casadaban MJ, Martinez-Arias A, Shapira SK, Chou J (1983) β-galactosidase gene fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol 100:293–308
Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156
Cooksey RC, Clark NC, Thornsberry C (1985) A gene probe for TEM type β-lactamases. Antimicrob Agents Chemother 28:154–156
Dale JW, Godwin D, Mossakowska D, Stephenson P, Wall S (1985) Sequence of the OXA2 β-lactamase: comparison with other penicillin-reactive enzymes. FEBS Lett 191:39–44
Grunstein M, Wallis J (1979) Colony hybridization. Methods Enzymol 68:379–389
Guerry P, LeBlanc DJ, Falkow S (1973) General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol 116:1064–1066
Hedges RW, Jacoby GA (1980) Compatibility and molecular properties of plasmid Rms 149 in Pseudomonas aeruginosa and Escherichia coli. Plasmid 3:1–6
Hedges RW, Medeiros AA, Cohenford M, Jacoby GA (1985) Genetic and biochemical properties of AER-1, a novel carbenicillin-hydrolyzing β-lactamase from Aeromonas hydrophila. Antimicrob Agents Chemother 27:479–484
Humphreys GO, Willshaw GA, Anderson ES (1975) A simple methods for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta 383:457–463
Jacob AE, Shapiro JA, Yamamoto L, Smith DI Cohen SN, Berg D (1977) Plasmids studied in Escherichia coli and other enteric bacteria. In: Buckhari AI, Shapiro JA, Adhya SL (eds) DNA insertion elements, plasmids, and episomes. Cold Spring Harbor Laboratory, New York 607–638
Jaurin B, Grundström T (1981) ampC cephalosporinase of Escherichia coli K-12 has a different evolutionary origin from that of β-lactamases of the penicillinase type. Proc Natl Acad Sci USA 78:4897–4901
Jaurin B, Grundström T, Normark S (1982) Sequence elements determining ampC promoter strength in E. coli. EMBO J 1:875–881
Kafatos FC, Jones CW, Efstratiadis A (1979) Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acid Res 7:1541–1552
Kahn M, Kolter R, Thomas C, Figurski D, Meyer R, Remaut E, Helinski DR (1979) Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol 68:268–280
Labia R, Guionie M, Barthélémy M, Philippon A (1981) Properties of three carbenicilin-hydrolyzing β-lactamases (CARB) from Pseudomonas aeruginosa: identification of a new enzyme. J Antimicrob Chemother 7:49–56
Lederberg EM, Cohen SN (1974) Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol 119:1072–1074
Levesque R, Roy PH (1982) Mapping of the plasmid (pLQ3) from Archromobacter and cloning of its cephalosporinase gene in Escherichia coli. Gene 18:69–75
Levesque R, Roy PH, Letarte R, Pechère J-C (1982) A plasmidmediated cephalosporinase from Achromobacter species. J Infect Dis 145:753–761
Levesque RC, Jacoby GA (1987) Molecular structure and interrelationships of multiresistance β-latamase transponsons (submitted for publication)
Livermore DM, Jones CS (1986) Characterization of NPS-1, a novel plasmid-mediated β-lactamase, from two Pseudomonas aeruginosa isolates. Antimicrob Agents Chemother 29:99–103
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, New York
Matthew M (1979) Plasmid-mediated β-lactamases of gram-negative bacteria: properties and distribution. J Antimicrob Chemother 5:349–358
Matthew M, Harris AM, Marshall MJ, Ross GW (1975) The use of analytical isoelectric focusing for detection and indentification of β-lactamases. J Gen Microbiol 88169–178
Matthew M, Hedges RW, Smith JT (1979) Types of β-lactamase determined by plasmids in gram-negative bacteria. J Bacteriol 138:657–662
Medeiros AA (1984) β-lactamases. Br Med Bull 40: 18–27
Medeiros AA, Cohenford M, Jacoby GA (1985) Five novel plasmid-determined β-lactamases. Antimicrob Agents chemother 27:715–719
Novick RP, Clowes RC, Cohen SN, Curtiss R, III, Data N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev 40:168–189
Philippon AM, Paul GC, Jacoby GA (1983) Properties of PSE-2 β-lactamase and genetic basis for its production in Pseudomonas aeruginosa. Antimicrob Agents Chemother 24:362–369
Philippon AM, Paul GC, Jacoby GA (1986a) New plasmid-mediated oxacillin-hydrolyzing β-lactamase in Pseudomonas aeruginosa. J Antimicrob Chemother 17:415–422
Philippon AM, Paul GC, Thabaut AP, Jacoby GA (1986b). Properties of a novel carbenicillin hydrolyzing β-lactamase (CARB-4) specified by an IncP-2 plasmid from Pseudomonas aeruginosa. Antimicrob Agents Chemother 29:519–520
Radloff R, Bauer W, Vinograd J (1967) A dye-boyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in Hela cells. Proc Natl Acad Sci USA 57:1514–1521
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Rubin LG, Medeiros AA, Yolken RH, Moxon ER (1981) Ampicillin treatment failure of apparently β-lactamase-negative Haemophilus influenzae type b meningitis due to novel β-lactamase. Lancet ii:1008–1010
Schroeder JL, Blattner FR (1978) Least-squares method for restriction mapping. Gene 4:167–174
Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Springer Harbor Laboratory, New York
Sutcliffe JG (1978) Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. Proc Natl Acad Sci USA 75:3737–3741
Sykes RB, Matthew M (1976) The β-lactamases of gram-negative bacteria. J Antimicrob Chemother 2:115–157
Ward JM, Grinsted J (1982) Physical and genetic analysis of the Inc-W group plasmids R388, Sa, and R7K. Plasmid 7:239–250
Author information
Authors and Affiliations
Additional information
Communicated by M. Takanami
Rights and permissions
About this article
Cite this article
Levesque, R.C., Medeiros, A.A. & Jacoby, G.A. Molecular cloning and DNA homology of plasmid-mediated β-lactamase genes. Mol Gen Genet 206, 252–258 (1987). https://doi.org/10.1007/BF00333581
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00333581