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. 1987 Apr;115(4):619–625. doi: 10.1093/genetics/115.4.619

Mode of Replicon Fusion Mediated by the Duplicated Insertion Sequence Is21 in Escherichia coli

Cornelia Reimmann 1, Dieter Haas 1
PMCID: PMC1203094  PMID: 3034717

Abstract

The insertion sequence IS21 (2.1 kb) originating from the broad-host-range IncP plasmid R68 transposes infrequently; by contrast, the IS21 tandem repeat found on the derivative R68.45 is highly active in transpositional mobilization of other replicons in a variety of Gram-negative bacteria. The mobilized plasmids are joined to R68.45 by single IS21 copies in direct orientation.—The formation of IS21 tandem duplications was observed in cointegrates between R68.45 and pBR325::IS21 and also in an RP1::IS21 plasmid derivative in which a segment located between two directly repeated copies of IS21 was deleted spontaneously. We speculate that IS21 tandem repeats can arise when the termini of two IS21 elements are specifically joined in a transposition or deletion event.—A resistance gene flanked by two IS21 elements in direct orientation did not behave as a transposon. The Ω fragment carrying transcription and translation stop signals was inserted into various sites of the IS21 tandem repeat; in this way it could be shown that the left IS21 element (which is next to the kanamycin resistance gene in R68.45) was 100 times more active in cointegrate formation than was the righthand element.—Cointegrates between the conjugative plasmid R751 and pBR325 derivatives carrying IS21 and IS21::Ω in tandem contained a single IS21 at one replicon junction and a single IS21::Ω at the other. In the IS21 duplications the inner IS21 ends were preferentially recognized (presumably by IS21 transposase), whereas the outer termini were not required for cointegrate formation. Based on these findings a conservative (simple) pathway of transposition is proposed for R68.45 and other plasmids with an IS 21 tandem repeat. In this model R68.45 is pictured as a large transposon whose ends are joined together to form a circular molecule which is capable of autonomous replication.

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Selected References

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

  1. Bennett P. M., Grinsted J., Richmond M. H. Transposition of TnA does not generate deletions. Mol Gen Genet. 1977 Jul 20;154(2):205–211. doi: 10.1007/BF00330839. [DOI] [PubMed] [Google Scholar]
  2. Danilevich V. N., Kostiuchenko D. A. Immunitet k povtornomu vstraivaniiu insertsionnoi posledovatel'nosti IS21. Mol Biol (Mosk) 1985 Sep-Oct;19(5):1242–1250. [PubMed] [Google Scholar]
  3. Depicker A., De Block M., Inzé D., Van Montagu M., Schell J. IS-like element IS8 in RP4 plasmid and its involvement in cointegration. Gene. 1980 Sep;10(4):329–338. doi: 10.1016/0378-1119(80)90153-5. [DOI] [PubMed] [Google Scholar]
  4. Grindley N. D., Reed R. R. Transpositional recombination in prokaryotes. Annu Rev Biochem. 1985;54:863–896. doi: 10.1146/annurev.bi.54.070185.004243. [DOI] [PubMed] [Google Scholar]
  5. Grinsted J., Bennett P. M., Higginson S., Richmond M. H. Regional preference of insertion of Tn501 and Tn802 into RP1 and its derivatives. Mol Gen Genet. 1978 Nov 9;166(3):313–320. doi: 10.1007/BF00267624. [DOI] [PubMed] [Google Scholar]
  6. Haas D., Riess G. Spontaneous deletions of the chromosome-mobilizing plasmid R68.45 in Pseudomonas aeruginosa PAO. Plasmid. 1983 Jan;9(1):42–52. doi: 10.1016/0147-619x(83)90030-6. [DOI] [PubMed] [Google Scholar]
  7. Hirsch P. R., Beringer J. E. A physical map of pPH1JI and pJB4JI. Plasmid. 1984 Sep;12(2):139–141. doi: 10.1016/0147-619x(84)90059-3. [DOI] [PubMed] [Google Scholar]
  8. Jeenes D. J., Soldati L., Baur H., Watson J. M., Mercenier A., Reimmann C., Leisinger T., Haas D. Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host. Mol Gen Genet. 1986 Jun;203(3):421–429. doi: 10.1007/BF00422066. [DOI] [PubMed] [Google Scholar]
  9. Morisato D., Kleckner N. Transposase promotes double strand breaks and single strand joints at Tn10 termini in vivo. Cell. 1984 Nov;39(1):181–190. doi: 10.1016/0092-8674(84)90204-6. [DOI] [PubMed] [Google Scholar]
  10. Pheiffer B. H., Zimmerman S. B. Polymer-stimulated ligation: enhanced blunt- or cohesive-end ligation of DNA or deoxyribooligonucleotides by T4 DNA ligase in polymer solutions. Nucleic Acids Res. 1983 Nov 25;11(22):7853–7871. doi: 10.1093/nar/11.22.7853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Riess G., Masepohl B., Puehler A. Analysis of IS21-mediated mobilization of plasmid pACYC184 by R68.45 in Escherichia coli. Plasmid. 1983 Sep;10(2):111–118. doi: 10.1016/0147-619x(83)90063-x. [DOI] [PubMed] [Google Scholar]
  12. Schurter W., Holloway B. W. Genetic analysis of promoters on the insertion sequence IS21 of plasmid R68.45. Plasmid. 1986 Jan;15(1):8–18. doi: 10.1016/0147-619x(86)90010-7. [DOI] [PubMed] [Google Scholar]
  13. Timmons M. S., Lieb M., Deonier R. C. Recombination between IS5 elements: requirement for homology and recombination functions. Genetics. 1986 Aug;113(4):797–810. doi: 10.1093/genetics/113.4.797. [DOI] [PMC free article] [PubMed] [Google Scholar]

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