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PMID: 10500193 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Hyperactive transposase mutants of the Himar1 mariner transposon.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 20 ·1999-09-28 ·Pages 11428-33

Lampe DJ, Akerley BJ, Rubin EJ, Mekalanos JJ, Robertson HM

Abstract

Mariner-family transposable elements are active in a wide variety of organisms and are becoming increasingly important genetic tools in species lacking sophisticated genetics. The Himar1 element, isolated from the horn fly, Haematobia irritans, is active in Escherichia coli when expressed appropriately. We used this fact to devise a genetic screen for hyperactive mutants of Himar1 transposase that enhance overall transposition from approximately 4- to 50-fold as measured in an E. coli assay. Purified mutant transposases retain their hyperactivity, although to a lesser degree, in an in vitro transposition assay. Mutants like those described herein should enable sophisticated analysis of the biochemistry of mariner transposition and should improve the use of these elements as genetic tools, both in vivo and in vitro.

MeSH Terms
Animals DNA/metabolism DNA Transposable Elements Muscidae/genetics Mutation Transposases/genetics
Chemicals
DNA Transposable Elements DNA Transposases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Lampe D J
Department of Biological Sciences, Duquesne University, Pittsburgh, PA 15282, USA. [email protected]
Akerley B J
Rubin E J
Mekalanos J J
Robertson H M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.20.11428
Email
[email protected]
Published
1999-09-28
Pages
11428-33
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC18050
Subset
IM
Grants
NIAID NIH HHS · R01 AI026289 · United States
NIAID NIH HHS · 2RO1AI33586-04 · United States
NIAID NIH HHS · AI-02137 · United States
NIAID NIH HHS · AI-26289 · United States
External Links
PubMed source DOI: 10.1073/pnas.96.20.11428 PMC Full Text View journal details
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