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PMID: 8220445 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

A versatile system for detecting transposition in Arabidopsis.

The Plant journal : for cell and molecular biology ·Vol. 3 ·No. 2 ·1993-02-00 ·Pages 273-89

Fedoroff NV, Smith DL

Abstract

The maize transposable element Activator (Ac) has been shown to be active in a number of dicots, including Arabidopsis thaliana, whose small genome and short generation time have favored its wide adoption as a model organism for molecular genetic approaches to plant physiology and development. Using the Ac element and several bacterial and plant marker genes, we have devised a versatile system for identifying plants in which a transposon has excised and reinserted elsewhere in the genome. The transposons have been designed to facilitate the identification of insertions downstream of promoters and in the vicinity of enhancers by the inclusion of a beta-glucuronidase (GUS) gene either lacking a promoter or having a minimal promoter sequence. The system permits the transposon and the source of transposase to be maintained either stably in separate plants or in the same plant. Plants in which transposition is occurring can be identified by the frequent somatic activation of the GUS gene. The herbicide chlorsulfuron is used as a selective agent to identify progeny plants in which the transposon has excised from its original insertion site within a chlorsulfuron-resistant acetolactate synthase gene. Additional selectable markers permit the identification of plants containing a transposed element, but lacking transposase. Here we describe our initial characterization of the system and demonstrate its reliability and efficiency in identifying plants with transposed elements.

MeSH Terms
Acetolactate Synthase/genetics Amidohydrolases/genetics Arabidopsis/drug effects,genetics Crosses, Genetic DNA Transposable Elements/genetics Gene Expression Regulation Genes, Reporter Genetic Markers Genetic Vectors/genetics Glucuronidase/biosynthesis,genetics Indoleacetic Acids/pharmacology Kanamycin Resistance/genetics Molecular Biology/methods Nucleotidyltransferases/genetics Plants, Genetically Modified Promoter Regions, Genetic/genetics Transformation, Genetic Transposases Zea mays/genetics
Chemicals
DNA Transposable Elements Genetic Markers Indoleacetic Acids indoleacetamide Acetolactate Synthase Nucleotidyltransferases Transposases Glucuronidase Amidohydrolases indoleacetamide hydrolase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fedoroff N V
Carnegie Institution of Washington, Department of Embryology, Baltimore, MD 21210.
Smith D L
Article Info
Journal
The Plant journal : for cell and molecular biology
Abbr.
Plant J
ISSN
0960-7412
Published
1993-02-00
Pages
273-89
Language
English
Region
England
NLM ID
9207397
Subset
IM
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