High rates of Ac/Ds germinal transposition in Arabidopsis suitable for gene isolation by insertional mutagenesis

Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6085-9. doi: 10.1073/pnas.89.13.6085.

Abstract

Overexpression of the Activator (Ac) transposase gene in Arabidopsis thaliana resulted in a minimal germinal transposition frequency of 27% in which independent Dissociation (Ds) transposition events were observed. Molecular analysis of 45 F1 generation Ac/Ds plants indicated that high rates of somatic excision had occurred, and independent germinal insertions were identified in F2 generation progeny plants. A tandem cauliflower mosaic virus (CaMV) promoter fused to two different Ac coding sequences significantly increased the rate of Ds transposition. The CaMV-Ac fusions activated single and multiple copies of two different Ds elements, DsDHFR and Ds35S-1, and reciprocal crosses resulted in similar transposition frequencies. The improved rate of independent germinal transposition observed makes Arabidopsis an ideal system for insertional mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Southern
  • Cloning, Molecular
  • DNA Transposable Elements*
  • Gene Expression
  • Genetic Vectors
  • Molecular Sequence Data
  • Mutagenesis, Insertional*
  • Nucleotidyltransferases / metabolism*
  • Oligodeoxyribonucleotides / chemistry
  • Plants / genetics*
  • RNA, Messenger / genetics
  • Rhizobium / genetics
  • Transposases
  • Zea mays / genetics

Substances

  • DNA Transposable Elements
  • Oligodeoxyribonucleotides
  • RNA, Messenger
  • Nucleotidyltransferases
  • Transposases