Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria

V de Lorenzo; M Herrero; U Jakubzik; K N Timmis

doi:10.1128/jb.172.11.6568-6572.1990

journal article Open Access Nov 01, 1990

Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria

V de Lorenzo M Herrero U Jakubzik K N Timmis

Journal of Bacteriology Vol. 172 No. 11 pp. 6568-6572 · American Society for Microbiology

View at Publisher Save 10.1128/jb.172.11.6568-6572.1990

Abstract

A collection of Tn5-derived minitransposons has been constructed that simplifies substantially the generation of insertion mutants, in vivo fusions with reporter genes, and the introduction of foreign DNA fragments into the chromosome of a variety of gram-negative bacteria, including the enteric bacteria and typical soil bacteria like Pseudomonas species. The minitransposons consist of genes specifying resistance to kanamycin, chloramphenicol, streptomycin-spectinomycin, and tetracycline as selection markers and a unique NotI cloning site flanked by 19-base-pair terminal repeat sequences of Tn5. Further derivatives also contain lacZ, phoA, luxAB, or xylE genes devoid of their native promoters located next to the terminal repeats in an orientation that affords the generation of gene-operon fusions. The transposons are located on a R6K-based suicide delivery plasmid that provides the IS50R transposase tnp gene in cis but external to the mobile element and whose conjugal transfer to recipients is mediated by RP4 mobilization functions in the donor.

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Details

Published: Nov 01, 1990
Vol/Issue: 172(11)
Pages: 6568-6572
License: View

Authors

V

V de Lorenzo

GBF-National Research Centre for Biotechnology, Braunschweig, Federal Republic of Germany.

M

M Herrero

GBF-National Research Centre for Biotechnology, Braunschweig, Federal Republic of Germany.

U

U Jakubzik

GBF-National Research Centre for Biotechnology, Braunschweig, Federal Republic of Germany.

K

K N Timmis

GBF-National Research Centre for Biotechnology, Braunschweig, Federal Republic of Germany.

Cite This Article

V de Lorenzo, M Herrero, U Jakubzik, et al. (1990). Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria. Journal of Bacteriology, 172(11), 6568-6572. https://doi.org/10.1128/jb.172.11.6568-6572.1990

Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria

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