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. 1981 Sep;147(3):851–859. doi: 10.1128/jb.147.3.851-859.1981

Tetracycline resistance transposon Tn1721: recA-dependent gene amplification and expression of tetracycline resistance.

K Wiebauer, S Schraml, S W Shales, R Schmitt
PMCID: PMC216121  PMID: 6268611

Abstract

The 7.1-megadalton transposon Tn1721 codes for inducible tetracycline resistance (Tcr). The transposable element consists of a "minor transposon" (3.6 megadaltons) encoding functions required for transposition and a "tet region" (3.5 megadaltons) encoding resistance. Multiple tandem repeats of the tet region can be generated by recA-dependent gene amplification. This feature of Tn1721 has been used to analyze the relationship between gene dosage and Tcr. Derivatives of plasmid R388:Tn1721 containing from one to nine copies of the tet region were isolated and separately transformed into recA host cells, where they are stably maintained. The results of the study of Tcr in these strains were as follows: (i) the uninduced, "basal" level of Tcr was linearly related to gene dosage between 4 and 36 copies of tet per chromosome equivalent; (ii) the underlying mechanism could not be attributed to reduced accumulation of the drug; and (iii) induction with tetracycline elicited a four- to fivefold reduction in drug accumulation, independent of the gene dosage.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altenbuchner J., Choi C. L., Grinsted J., Schmitt R., Richmond M. H. The transposons Tn501(Hg) and Tn1721(Tc) are related. Genet Res. 1981 Jun;37(3):285–289. doi: 10.1017/s0016672300020280. [DOI] [PubMed] [Google Scholar]
  2. Anderson R. P., Roth J. R. Tandem genetic duplications in Salmonella typhimurium: amplification of the histidine operon. J Mol Biol. 1978 Nov 25;126(1):53–71. doi: 10.1016/0022-2836(78)90279-6. [DOI] [PubMed] [Google Scholar]
  3. Appleyard R K. Segregation of New Lysogenic Types during Growth of a Doubly Lysogenic Strain Derived from Escherichia Coli K12. Genetics. 1954 Jul;39(4):440–452. doi: 10.1093/genetics/39.4.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ball P. R., Shales S. W., Chopra I. Plasmid-mediated tetracycline resistance in Escherichia coli involves increased efflux of the antibiotic. Biochem Biophys Res Commun. 1980 Mar 13;93(1):74–81. doi: 10.1016/s0006-291x(80)80247-6. [DOI] [PubMed] [Google Scholar]
  6. Barbour S. D., Nagaishi H., Templin A., Clark A. J. Biochemical and genetic studies of recombination proficiency in Escherichia coli. II. Rec+ revertants caused by indirect suppression of rec- mutations. Proc Natl Acad Sci U S A. 1970 Sep;67(1):128–135. doi: 10.1073/pnas.67.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Beck C. F. A genetic approach to analysis of transposons. Proc Natl Acad Sci U S A. 1979 May;76(5):2376–2380. doi: 10.1073/pnas.76.5.2376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Boyer H. W., 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. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  9. Burkardt H. J., Mattes R., Schmid K., Schmitt R. Properties of two conjugative plasmids mediating tetracycline resistance, raffinose catabolism and hydrogen sulfide production in Escherichia coli. Mol Gen Genet. 1978 Oct 25;166(1):75–84. doi: 10.1007/BF00379731. [DOI] [PubMed] [Google Scholar]
  10. Cabello F., Timmis K., Cohen S. N. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature. 1976 Jan 29;259(5541):285–290. doi: 10.1038/259285a0. [DOI] [PubMed] [Google Scholar]
  11. Chabbert Y. A., Scavizzi M. R. Chelocardin-inducible resistance in Escherichia coli bearing R plasmids. Antimicrob Agents Chemother. 1976 Jan;9(1):36–41. doi: 10.1128/aac.9.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chandler M., de la Tour E. B., Willems D., Caro L. Some properties of the chloramphenicol resistance transposon Tn9. Mol Gen Genet. 1979 Oct 3;176(2):221–231. doi: 10.1007/BF00273216. [DOI] [PubMed] [Google Scholar]
  13. Chopra I., Howe T. G. Bacterial resistance to the tetracyclines. Microbiol Rev. 1978 Dec;42(4):707–724. doi: 10.1128/mr.42.4.707-724.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Datta N., Hedges R. W. Trimethoprim resistance conferred by W plasmids in Enterobacteriaceae. J Gen Microbiol. 1972 Sep;72(2):349–355. doi: 10.1099/00221287-72-2-349. [DOI] [PubMed] [Google Scholar]
  16. Hashimoto H., Rownd R. H. Transition of the R factor NR1 and Proteus mirabilis: level of drug resistance of nontransitioned and transitioned cells. J Bacteriol. 1975 Jul;123(1):56–68. doi: 10.1128/jb.123.1.56-68.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
  18. Izaki K., Kiuchi K., Arima K. Specificity and mechanism of tetracycline resistance in a multiple drug resistant strain of Escherichia coli. J Bacteriol. 1966 Feb;91(2):628–633. doi: 10.1128/jb.91.2.628-633.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jorgensen R. A., Reznikoff W. S. Organization of structural and regulatory genes that mediate tetracycline resistance in transposon Tn10. J Bacteriol. 1979 Jun;138(3):705–714. doi: 10.1128/jb.138.3.705-714.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Levy S. B., McMurry L. Plasmid-determined tetracycline resistance involves new transport systems for tetracycline. Nature. 1978 Nov 2;276(5683):90–92. doi: 10.1038/276090a0. [DOI] [PubMed] [Google Scholar]
  21. Maloney P. C., Rotman B. Distribution of suboptimally induces -D-galactosidase in Escherichia coli. The enzyme content of individual cells. J Mol Biol. 1973 Jan;73(1):77–91. doi: 10.1016/0022-2836(73)90160-5. [DOI] [PubMed] [Google Scholar]
  22. Mattes R., Burkardt H. J., Schmitt R. Repetition of tetracycline resistance determinant genes on R plasmid pRSD1 in Escherichia coli. Mol Gen Genet. 1979 Jan 10;168(2):173–184. doi: 10.1007/BF00431443. [DOI] [PubMed] [Google Scholar]
  23. McMurry L., Petrucci R. E., Jr, Levy S. B. Active efflux of tetracycline encoded by four genetically different tetracycline resistance determinants in Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3974–3977. doi: 10.1073/pnas.77.7.3974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mendez B., Tachibana C., Levy S. B. Heterogeneity of tetracycline resistance determinants. Plasmid. 1980 Mar;3(2):99–108. doi: 10.1016/0147-619x(80)90101-8. [DOI] [PubMed] [Google Scholar]
  25. Meyer J., Iida S. Amplification of chloramphenicol resistance transposons carried by phage P1Cm in Escherichia coli. Mol Gen Genet. 1979 Oct 3;176(2):209–219. doi: 10.1007/BF00273215. [DOI] [PubMed] [Google Scholar]
  26. Nierhaus K. H., Wittmann H. G. Ribosomal function and its inhibition by antibiotics in prokaryotes. Naturwissenschaften. 1980 May;67(5):234–250. doi: 10.1007/BF01054532. [DOI] [PubMed] [Google Scholar]
  27. Normark S., Edlund T., Grundström T., Bergström S., Wolf-Watz H. Escherichia coli K-12 mutants hyperproducing chromosomal beta-lactamase by gene repetitions. J Bacteriol. 1977 Dec;132(3):912–922. doi: 10.1128/jb.132.3.912-922.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Novick R. P., Clowes R. C., Cohen S. N., Curtiss R., 3rd, Datta N., Falkow S. Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev. 1976 Mar;40(1):168–189. doi: 10.1128/br.40.1.168-189.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rownd R., Mickel S. Dissociation and reassociation of RTF and r-determinants of the R-factor NR1 in Proteus mirabilis. Nat New Biol. 1971 Nov 10;234(45):40–43. doi: 10.1038/newbio234040a0. [DOI] [PubMed] [Google Scholar]
  30. Schmitt R., Altenbuchner J., Wiebauer K., Arnold W., Pühler A., Schöffl F. Basis of transposition and gene amplification by Tn1721 and related tetracycline-resistance transposons. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):59–65. doi: 10.1101/sqb.1981.045.01.011. [DOI] [PubMed] [Google Scholar]
  31. Schmitt R., Bernhard E., Mattes R. Characterisation of Tn1721, a new transposon containing tetracycline resistance genes capable of amplification. Mol Gen Genet. 1979 Apr 17;172(1):53–65. doi: 10.1007/BF00276215. [DOI] [PubMed] [Google Scholar]
  32. Schöffl F., Arnold W., Pühler A., Altenbuchner J., Schmitt R. The tetracycline resistance transposons Tn1721 and Tn1771 have three 38-base-pair repeats and generate five-base-pair direct repeats. Mol Gen Genet. 1981;181(1):87–94. doi: 10.1007/BF00339010. [DOI] [PubMed] [Google Scholar]
  33. Schöffl F., Pühler A. Intramolecular amplification of the tetracycline resistance determinant of transposon Tn1771 in Escherichia coli. Genet Res. 1979 Jun;33(3):253–260. doi: 10.1017/s0016672300018395. [DOI] [PubMed] [Google Scholar]
  34. Shales S. W., Chopra I., Ball P. R. Evidence for more than one mechanism of plasmid-determined tetracycline resistance in Escherichia coli. J Gen Microbiol. 1980 Nov;121(1):221–229. doi: 10.1099/00221287-121-1-221. [DOI] [PubMed] [Google Scholar]
  35. Tait R. C., Boyer H. W. On the nature of tetracycline resistance controlled by the plasmid pSC101. Cell. 1978 Jan;13(1):73–81. doi: 10.1016/0092-8674(78)90139-3. [DOI] [PubMed] [Google Scholar]
  36. Tanaka T., Weisblum B. Construction of a colicin E1-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid. J Bacteriol. 1975 Jan;121(1):354–362. doi: 10.1128/jb.121.1.354-362.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Taylor D. P., Greenberg J., Rownd R. H. Generation of miniplasmids from copy number mutants of the R plasmid NR1. J Bacteriol. 1977 Dec;132(3):986–995. doi: 10.1128/jb.132.3.986-995.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ward J. M., Grinsted J. Mapping of functions in the R-plasmid R388 by examination of deletion mutants generated in vitro. Gene. 1978 Apr;3(2):87–95. doi: 10.1016/0378-1119(78)90053-7. [DOI] [PubMed] [Google Scholar]
  39. Yagi Y., Clewell D. B. Amplification of the tetracycline resistance determinant of plasmid pAM alpha 1 in Streptococcus faecalis: dependence on host recombination machinery. J Bacteriol. 1980 Aug;143(2):1070–1072. doi: 10.1128/jb.143.2.1070-1072.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yang H. L., Zubay G., Levy S. B. Synthesis of an R plasmid protein associated with tetracycline resistance is negatively regulated. Proc Natl Acad Sci U S A. 1976 May;73(5):1509–1512. doi: 10.1073/pnas.73.5.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zupancic T. J., King S. R., Pogue-Geile K. L., Jaskunas S. R. Identification of a second tetracycline-inducible polypeptide encoded by Tn10. J Bacteriol. 1980 Oct;144(1):346–355. doi: 10.1128/jb.144.1.346-355.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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