Resources for targeted insertional and deletional mutagenesis in Arabidopsis

Sen Zhang, Surabhi Raina, Hong Li, Jun Li, Ewa Dec, Hong Ma, Hai Huang, Nina V. Fedoroff

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The maize transposons Activator (Ac) and Dissociation (Ds) are active in many monocots and dicots, including Arabidopsis. We describe a new Ac-derived transposon construct, designated the Ds-loxP T-DNA, which can be used for both insertional and deletional mutagenesis. There are loxP sites in both orientations on both the transposon and the donor site T-DNA and an arrangement of marker genes that permits selection of transposition events, as well as deletions and inversions extending from the donor site to a transposon reinserted on either side of it. We show that Cre-mediated deletions and inversions occur at a high frequency. The tendency of Ac-Ds transposons to reinsert near the donor site can be used to target both insertional and deletional mutagenesis, but efficient exploitation of this property requires a library of mapped marked donor sites distributed in the genome. We have created a population of independent Ds T-DNA transformants and we have mapped an initial set of 75 Ds T-DNA integration sites. We assessed the potential efficiency of targeted mutagenesis by detecting Ds reinsertion events at several loci over a 400 kb interval from each of two donor sites with different Ds T-DNA constructs. The distribution of reinsertion sites is similar around the two tested loci, with roughly 10, 4, and ca. 1% of reinsertions detected within 1-2 kb of sites 10, 100, and 200-400 kb from the donor site, respectively. To facilitate the use of this targeted mutagenesis system, we have constructed a searchable database of the mapped Ds T-DNA integration sites.

Original languageEnglish (US)
Pages (from-to)133-150
Number of pages18
JournalPlant molecular biology
Volume53
Issue number1-2
DOIs
StatePublished - Sep 1 2003

Fingerprint

Insertional Mutagenesis
Arabidopsis
mutagenesis
transposons
DNA
Mutagenesis
site-directed mutagenesis
Gene Order
Genetic Markers
loci
Libraries
Zea mays
T-DNA
transposition (genetics)
Magnoliopsida
Liliopsida
Genome
Databases
Population
genetic markers

All Science Journal Classification (ASJC) codes

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

Cite this

Zhang, Sen ; Raina, Surabhi ; Li, Hong ; Li, Jun ; Dec, Ewa ; Ma, Hong ; Huang, Hai ; Fedoroff, Nina V. / Resources for targeted insertional and deletional mutagenesis in Arabidopsis. In: Plant molecular biology. 2003 ; Vol. 53, No. 1-2. pp. 133-150.
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Zhang, S, Raina, S, Li, H, Li, J, Dec, E, Ma, H, Huang, H & Fedoroff, NV 2003, 'Resources for targeted insertional and deletional mutagenesis in Arabidopsis', Plant molecular biology, vol. 53, no. 1-2, pp. 133-150. https://doi.org/10.1023/B:PLAN.0000009271.08420.d9

Resources for targeted insertional and deletional mutagenesis in Arabidopsis. / Zhang, Sen; Raina, Surabhi; Li, Hong; Li, Jun; Dec, Ewa; Ma, Hong; Huang, Hai; Fedoroff, Nina V.

In: Plant molecular biology, Vol. 53, No. 1-2, 01.09.2003, p. 133-150.

Research output: Contribution to journalArticle

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