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Related Experiment Videos

Mapping Ds insertions in barley using a sequence-based approach.

L D Cooper1, L Marquez-Cedillo, J Singh

  • 1Department of Crop and Soil Science, Oregon State University, OR 97331, Corvallis, USA.

Molecular Genetics and Genomics : MGG
|September 28, 2004
PubMed
Summary
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A new transposon tagging system using maize Ac/Ds elements was developed in barley. This system facilitates gene discovery and reverse genetics by creating Ds insertion lines throughout the barley genome.

Area of Science:

  • Plant genetics
  • Molecular biology
  • Genomics

Background:

  • Maize Ac/Ds elements offer a robust system for transposon tagging.
  • Developing such a system in barley is crucial for functional genomics in Triticeae.

Purpose of the Study:

  • To establish a transposon tagging system in barley using maize Ac/Ds elements.
  • To generate a collection of barley lines with Ds insertions dispersed across the genome for gene discovery.

Main Methods:

  • Biolistic transformation of barley with AcTPase and Ds-bar elements.
  • Inverse PCR and TAIL-PCR for flanking sequence identification.
  • Sequence-based mapping using SNP and indel polymorphisms in the Oregon Wolfe Barley population.

Main Results:

Related Experiment Videos

  • 19 independent barley lines with stabilized Ds insertions at new genomic locations were generated.
  • Genome locations of Ds insertions were determined using sequence-based mapping and confirmed with RFLP analysis.
  • BLAST searches revealed that most Ds insertions are within known or predicted coding sequences.

Conclusions:

  • The Ac/Ds transposon tagging system is effective in barley.
  • This system provides a valuable tool for functional genomics and gene discovery in barley and other Triticeae species.