Transposon Tn5 target specificity: preference for insertion at G/C pairs.

Journal Article (Journal Article)

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

Duke Authors

Cited Authors

  • Lodge, JK; Weston-Hafer, K; Berg, DE

Published Date

  • November 1988

Published In

Volume / Issue

  • 120 / 3

Start / End Page

  • 645 - 650

PubMed ID

  • 2852135

Pubmed Central ID

  • PMC1203542

International Standard Serial Number (ISSN)

  • 0016-6731

Digital Object Identifier (DOI)

  • 10.1093/genetics/120.3.645


  • eng

Conference Location

  • United States