Large Scale Technologies and Resources
Singh, Jaswinder , Vutien, Philip , Blosse, Pierre-Alain , Gong, Alice , Sandhu, Harjot , Shafir, Adi , Sotelo-Troha, Katia , Lemaux, Peggy G. .
Transposon-mediated Gene Search in Barley.
The maize Ac/Ds transposon system is an effective approach for gene identification and cloning in heterologous species. Using this system, single-copy Ds insertion lines (TNPs) were generated in barley to identify and tag genes and conduct gene function studies. With the availability of extensive genomics resources in barley, a robust platform is available for the effective use of the transposon approaches to characterizing and isolating genes. From 200 independent TNPs, flanking sequences adjacent to Ds were determined in >100 lines. Results from BLAST searches indicate ~86% of Ds flanking sequences are from known or putative genes, which encode, for example, MLA1, wall-associated kinases, ubiquitin-conjugating enzyme, ATP-binding transporter, terpene synthase, ankyrin 1 and cytochrome P450. As mapping information on these insertion sites evolves, that information can lead to strategies to tag agronomically important genes by identifying Ds elements that are in close proximity to traits/genes of interest, followed by Ds reactivation to achieve saturation mutagenesis. For maximal utility of this approach, we undertook a study to understand the characteristics of TNPs important to reactivation in barley, i.e., status of terminal inverted repeats (TIRs) and 8 bp duplications and the nature of insertion sites. In addition, TNP re-activation frequencies over 3-4 consecutive generations of transposition were determined to provide the foundation for tagging and “transposon-walking” approaches.
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1 - University of California, Berkeley
2 - University of California, Berkeley, Plant and Microbial Biology, 111 Koshland Hall, Berkeley, CA, 94720-3102, USA
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM