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. 1995 Jan;39(1):155–162. doi: 10.1128/aac.39.1.155

Expression of antibiotic resistance genes in the integrated cassettes of integrons.

C M Collis 1, R M Hall 1
PMCID: PMC162502  PMID: 7695299

Abstract

Plasmids containing cloned integron fragments which differ only with respect to either the sequence of the promoter(s) or the number and order of inserted cassettes were used to examine the expression of resistance genes encoded in integron-associated gene cassettes. All transcripts detected commenced at the common promoter P(ant), and alterations in the sequence of P(ant) affected the level of resistance expressed by cassette genes. When both P(ant) and the secondary promoter P2 were present, transcription from both promoters was detected. When more than one cassette was present, the position of the cassette in the array influenced the level of antibiotic resistance expressed by the cassette gene. In all cases, the resistance level was highest when the gene was in the first cassette, i.e., closest to P(ant), and was reduced to different extents by the presence of individual upstream cassettes. In Northern (RNA) blots, multiple discrete transcripts originating at P(ant) were detected, and only the longer transcripts contained the distal genes. Together, these data suggest that premature transcription termination occurs within the cassettes. The most abundant transcripts appeared to contain one or more complete cassettes, and is possible that the 59-base elements found at the end of the cassettes (3' to the coding region) not only function as recombination sites but may also function as transcription terminators.

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Selected References

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

  1. Belasco J. G., Higgins C. F. Mechanisms of mRNA decay in bacteria: a perspective. Gene. 1988 Dec 10;72(1-2):15–23. doi: 10.1016/0378-1119(88)90123-0. [DOI] [PubMed] [Google Scholar]
  2. Bissonnette L., Champetier S., Buisson J. P., Roy P. H. Characterization of the nonenzymatic chloramphenicol resistance (cmlA) gene of the In4 integron of Tn1696: similarity of the product to transmembrane transport proteins. J Bacteriol. 1991 Jul;173(14):4493–4502. doi: 10.1128/jb.173.14.4493-4502.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bito A., Susani M. Revised analysis of aadA2 gene of plasmid pSa. Antimicrob Agents Chemother. 1994 May;38(5):1172–1175. doi: 10.1128/aac.38.5.1172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cameron F. H., Groot Obbink D. J., Ackerman V. P., Hall R. M. Nucleotide sequence of the AAD(2'') aminoglycoside adenylyltransferase determinant aadB. Evolutionary relationship of this region with those surrounding aadA in R538-1 and dhfrII in R388. Nucleic Acids Res. 1986 Nov 11;14(21):8625–8635. doi: 10.1093/nar/14.21.8625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Collis C. M., Grammaticopoulos G., Briton J., Stokes H. W., Hall R. M. Site-specific insertion of gene cassettes into integrons. Mol Microbiol. 1993 Jul;9(1):41–52. doi: 10.1111/j.1365-2958.1993.tb01667.x. [DOI] [PubMed] [Google Scholar]
  7. Collis C. M., Hall R. M. Gene cassettes from the insert region of integrons are excised as covalently closed circles. Mol Microbiol. 1992 Oct;6(19):2875–2885. doi: 10.1111/j.1365-2958.1992.tb01467.x. [DOI] [PubMed] [Google Scholar]
  8. Collis C. M., Hall R. M. Site-specific deletion and rearrangement of integron insert genes catalyzed by the integron DNA integrase. J Bacteriol. 1992 Mar;174(5):1574–1585. doi: 10.1128/jb.174.5.1574-1585.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hall R. M., Brookes D. E., Stokes H. W. Site-specific insertion of genes into integrons: role of the 59-base element and determination of the recombination cross-over point. Mol Microbiol. 1991 Aug;5(8):1941–1959. doi: 10.1111/j.1365-2958.1991.tb00817.x. [DOI] [PubMed] [Google Scholar]
  10. Hall R. M., Vockler C. The region of the IncN plasmid R46 coding for resistance to beta-lactam antibiotics, streptomycin/spectinomycin and sulphonamides is closely related to antibiotic resistance segments found in IncW plasmids and in Tn21-like transposons. Nucleic Acids Res. 1987 Sep 25;15(18):7491–7501. doi: 10.1093/nar/15.18.7491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hollingshead S., Vapnek D. Nucleotide sequence analysis of a gene encoding a streptomycin/spectinomycin adenylyltransferase. Plasmid. 1985 Jan;13(1):17–30. doi: 10.1016/0147-619x(85)90052-6. [DOI] [PubMed] [Google Scholar]
  12. Lévesque C., Brassard S., Lapointe J., Roy P. H. Diversity and relative strength of tandem promoters for the antibiotic-resistance genes of several integrons. Gene. 1994 May 3;142(1):49–54. doi: 10.1016/0378-1119(94)90353-0. [DOI] [PubMed] [Google Scholar]
  13. Mossakowska D., Ali N. A., Dale J. W. Oxacillin-hydrolysing beta-lactamases. A comparative analysis at nucleotide and amino acid sequence levels. Eur J Biochem. 1989 Mar 15;180(2):309–318. doi: 10.1111/j.1432-1033.1989.tb14649.x. [DOI] [PubMed] [Google Scholar]
  14. Newbury S. F., Smith N. H., Higgins C. F. Differential mRNA stability controls relative gene expression within a polycistronic operon. Cell. 1987 Dec 24;51(6):1131–1143. doi: 10.1016/0092-8674(87)90599-x. [DOI] [PubMed] [Google Scholar]
  15. Recchia G. D., Stokes H. W., Hall R. M. Characterisation of specific and secondary recombination sites recognised by the integron DNA integrase. Nucleic Acids Res. 1994 Jun 11;22(11):2071–2078. doi: 10.1093/nar/22.11.2071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schmidt F. R., Nücken E. J., Henschke R. B. Nucleotide sequence analysis of 2''-aminoglycoside nucleotidyl-transferase ANT(2'') from Tn4000: its relationship with AAD(3'') and impact on Tn21 evolution. Mol Microbiol. 1988 Nov;2(6):709–717. doi: 10.1111/j.1365-2958.1988.tb00081.x. [DOI] [PubMed] [Google Scholar]
  17. Stephen D., Jones C., Schofield J. P. A rapid method for isolating high quality plasmid DNA suitable for DNA sequencing. Nucleic Acids Res. 1990 Dec 25;18(24):7463–7464. doi: 10.1093/nar/18.24.7463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stokes H. W., Hall R. M. A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: integrons. Mol Microbiol. 1989 Dec;3(12):1669–1683. doi: 10.1111/j.1365-2958.1989.tb00153.x. [DOI] [PubMed] [Google Scholar]
  19. Stokes H. W., Hall R. M. Sequence analysis of the inducible chloramphenicol resistance determinant in the Tn1696 integron suggests regulation by translational attenuation. Plasmid. 1991 Jul;26(1):10–19. doi: 10.1016/0147-619x(91)90032-r. [DOI] [PubMed] [Google Scholar]
  20. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Summers W. C. A simple method for extraction of RNA from E. coli utilizing diethyl pyrocarbonate. Anal Biochem. 1970 Feb;33(2):459–463. doi: 10.1016/0003-2697(70)90316-7. [DOI] [PubMed] [Google Scholar]
  22. Swift G., McCarthy B. J., Heffron F. DNA sequence of a plasmid-encoded dihydrofolate reductase. Mol Gen Genet. 1981;181(4):441–447. doi: 10.1007/BF00428733. [DOI] [PubMed] [Google Scholar]
  23. Tait R. C., Rempel H., Rodriguez R. L., Kado C. I. The aminoglycoside-resistance operon of the plasmid pSa: nucleotide sequence of the streptomycin-spectinomycin resistance gene. Gene. 1985;36(1-2):97–104. doi: 10.1016/0378-1119(85)90073-3. [DOI] [PubMed] [Google Scholar]
  24. Tenover F. C., Filpula D., Phillips K. L., Plorde J. J. Cloning and sequencing of a gene encoding an aminoglycoside 6'-N-acetyltransferase from an R factor of Citrobacter diversus. J Bacteriol. 1988 Jan;170(1):471–473. doi: 10.1128/jb.170.1.471-473.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wohlleben W., Arnold W., Bissonnette L., Pelletier A., Tanguay A., Roy P. H., Gamboa G. C., Barry G. F., Aubert E., Davies J. On the evolution of Tn21-like multiresistance transposons: sequence analysis of the gene (aacC1) for gentamicin acetyltransferase-3-I(AAC(3)-I), another member of the Tn21-based expression cassette. Mol Gen Genet. 1989 Jun;217(2-3):202–208. doi: 10.1007/BF02464882. [DOI] [PubMed] [Google Scholar]
  26. de la Cruz F., Grinsted J. Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1. J Bacteriol. 1982 Jul;151(1):222–228. doi: 10.1128/jb.151.1.222-228.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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