• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of geneticsGeneticsCurrent IssueInformation for AuthorsEditorial BoardSubscribeSubmit a Manuscript
Genetics. Nov 1988; 120(3): 645–650.
PMCID: PMC1203542

Transposon Tn5 Target Specificity: Preference for Insertion at G/C Pairs


The procaryotic transposon Tn5 inserts into many different sites within a single gene, but some sites (hotspots) are targeted repeatedly. Hotspots are not closely related in sequence, but most have G/C pairs at the ends of the nine base pairs duplicated by Tn5 insertion. In pBR322, the major hotspot coincides with the ``-10 region'' of the tet promoter. We mutated the G/C pairs at this hotspot and assayed for insertion into hotspot I, resistance to tetracycline, and plasmid supercoiling. We found that changing the G/C pairs to A/T pairs reduced the frequency of insertion into the hotspot by at least fivefold. The reduction in hotspot use caused by these G/C to A/T changes was not attributable to changes in plasmid supercoiling or tet promoter strength.

Full Text

The Full Text of this article is available as a PDF (2.5M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Berg DE, Schmandt MA, Lowe JB. Specificity of transposon Tn5 insertion. Genetics. 1983 Dec;105(4):813–828. [PMC free article] [PubMed]
  • Brosius J, Cate RL, Perlmutter AP. Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro. J Biol Chem. 1982 Aug 10;257(15):9205–9210. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Collins J, Volckaert G, Nevers P. Precise and nearly-precise excision of the symmetrical inverted repeats of Tn5; common features of recA-independent deletion events in Escherichia coli. Gene. 1982 Jul-Aug;19(1):139–146. [PubMed]
  • Craigie R, Arndt-Jovin DJ, Mizuuchi K. A defined system for the DNA strand-transfer reaction at the initiation of bacteriophage Mu transposition: protein and DNA substrate requirements. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7570–7574. [PMC free article] [PubMed]
  • Craigie R, Mizuuchi K. Mechanism of transposition of bacteriophage Mu: structure of a transposition intermediate. Cell. 1985 Jul;41(3):867–876. [PubMed]
  • Dewitt SK, Adelberg EA. The Occurrence of a Genetic Transposition in a Strain of Escherichia Coli. Genetics. 1962 May;47(5):577–585. [PMC free article] [PubMed]
  • Egelhoff TT, Fisher RF, Jacobs TW, Mulligan JT, Long SR. Nucleotide sequence of Rhizobium meliloti 1021 nodulation genes: nodD is read divergently from nodABC. DNA. 1985 Jun;4(3):241–248. [PubMed]
  • Galas DJ, Calos MP, Miller JH. Sequence analysis of Tn9 insertions in the lacZ gene. J Mol Biol. 1980 Nov 25;144(1):19–41. [PubMed]
  • Gamas P, Chandler MG, Prentki P, Galas DJ. Escherichia coli integration host factor binds specifically to the ends of the insertion sequence IS1 and to its major insertion hot-spot in pBR322. J Mol Biol. 1987 May 20;195(2):261–272. [PubMed]
  • Halling SM, Kleckner N. A symmetrical six-base-pair target site sequence determines Tn10 insertion specificity. Cell. 1982 Jan;28(1):155–163. [PubMed]
  • Klaer R, Kühn S, Fritz HJ, Tillmann E, Saint-Girons I, Habermann P, Pfeifer D, Starlinger P. Studies on transposition mechanisms and specificity of IS4. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):215–224. [PubMed]
  • Isberg RR, Syvanen M. DNA gyrase is a host factor required for transposition of Tn5. Cell. 1982 Aug;30(1):9–18. [PubMed]
  • Lupski JR, Gershon P, Ozaki LS, Godson GN. Specificity of Tn5 insertions into a 36-bp DNA sequence repeated in tandem seven times. Gene. 1984 Oct;30(1-3):99–106. [PubMed]
  • Mandecki W. Oligonucleotide-directed double-strand break repair in plasmids of Escherichia coli: a method for site-specific mutagenesis. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7177–7181. [PMC free article] [PubMed]
  • McKown RL, Waddell CS, Arciszewska LK, Craig NL. Identification of a transposon Tn7-dependent DNA-binding activity that recognizes the ends of Tn7. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7807–7811. [PMC free article] [PubMed]
  • Peden KW. Revised sequence of the tetracycline-resistance gene of pBR322. Gene. 1983 May-Jun;22(2-3):277–280. [PubMed]
  • Pruss GJ. DNA topoisomerase I mutants. Increased heterogeneity in linking number and other replicon-dependent changes in DNA supercoiling. J Mol Biol. 1985 Sep 5;185(1):51–63. [PubMed]
  • Schaller H. The intergenic region and the origins for filamentous phage DNA replication. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):401–408. [PubMed]
  • Sengstag C, Arber W. A cloned DNA fragment from bacteriophage P1 enhances IS2 insertion. Mol Gen Genet. 1987 Feb;206(2):344–351. [PubMed]
  • Shortle D, Nathans D. Local mutagenesis: a method for generating viral mutants with base substitutions in preselected regions of the viral genome. Proc Natl Acad Sci U S A. 1978 May;75(5):2170–2174. [PMC free article] [PubMed]
  • Sternglanz R, DiNardo S, Voelkel KA, Nishimura Y, Hirota Y, Becherer K, Zumstein L, Wang JC. Mutations in the gene coding for Escherichia coli DNA topoisomerase I affect transcription and transposition. Proc Natl Acad Sci U S A. 1981 May;78(5):2747–2751. [PMC free article] [PubMed]
  • Noll M. Internal structure of the nucleosome: DNA folding in the conserved 140-base-pair core particle. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):77–85. [PubMed]
  • Syvanen M, Hopkins JD, Clements M. A new class of mutants in DNA polymerase I that affects gene transposition. J Mol Biol. 1982 Jun 25;158(2):203–212. [PubMed]
  • Tu CP, Cohen SN. Translocation specificity of the Tn3 element: characterization of sites of multiple insertions. Cell. 1980 Jan;19(1):151–160. [PubMed]
  • Waddell CS, Craig NL. Tn7 transposition: two transposition pathways directed by five Tn7-encoded genes. Genes Dev. 1988 Feb;2(2):137–149. [PubMed]
  • Yin JC, Reznikoff WS. dnaA, an essential host gene, and Tn5 transposition. J Bacteriol. 1987 Oct;169(10):4637–4645. [PMC free article] [PubMed]
  • Zerbib D, Gamas P, Chandler M, Prentki P, Bass S, Galas D. Specificity of insertion of IS1. J Mol Biol. 1985 Oct 5;185(3):517–524. [PubMed]
  • Zoller MJ, Smith M. Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA. Nucleic Acids Res. 1982 Oct 25;10(20):6487–6500. [PMC free article] [PubMed]

Articles from Genetics are provided here courtesy of Genetics Society of America


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...