Sequence requirements for target activity in site‐specific recombination mediated by the Int protein of transposon Tn 1545
- 1 April 1993
- journal article
- Published by Wiley in Molecular Microbiology
- Vol. 8 (1), 179-185
- https://doi.org/10.1111/j.1365-2958.1993.tb01214.x
Abstract
Excision and integration of Tn 1545 occur by reciprocal site‐specific recombination between 6 (or 7)bp variable sequences present in the recombining attachment (atf) sites and designated overlap regions. We devised an assay for Tn1545 transposition in which derivatives containing the cis‐acting transposition sequences (atfTn 1545) integrate into a target replicon when complemented in trans by the transposon‐encoded integrase Int‐Tn. This assay was used to determine the characteristics of the DNA sequence that influence target site selection. Characterization of several integration sites indicated that a 20 bp segment, designated attB, contains the sequences required for target activity. It also appeared that (i) the target activity depends upon the extent of homology between the 7bp segments flanking the overlap regions in attB and attTn1545, and (ii) the degree of homology between the two recombining overlap regions does not affect the level of target activity and has no influence on integration orientation.Keywords
This publication has 21 references indexed in Scilit:
- An integrative vector exploiting the transposition properties of Tn1545 for insertional mutagenesis and cloning of genes from Gram-positive bacteriaGene, 1991
- Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from gram-positive cocciCell, 1991
- The integration‐excision system of the conjugative transposon Tn 1545 is structurally and functionally related to those of lambdoid phagesMolecular Microbiology, 1990
- Genetic analysis of a tetracycline resistance element from Clostridium difficile and its conjugal transfer to and from Bacillus subtilisJournal of General Microbiology, 1990
- Excision and insertion of the conjugative transposon Tn916 involves a novel recombination mechanismCell, 1989
- Transfer of Tn1545 and Tn916 to Clostridium acetobutylicumPlasmid, 1989
- DYNAMIC, STRUCTURAL, AND REGULATORY ASPECTS OF λ SITE-SPECIFIC RECOMBINATIONAnnual Review of Biochemistry, 1989
- Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectorsGene, 1985
- A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectorsGene, 1984
- A Broad Host Range Mobilization System for In Vivo Genetic Engineering: Transposon Mutagenesis in Gram Negative BacteriaBio/Technology, 1983