NDLI: High-frequency Ds remobilization over multiple generations in barley facilitates gene tagging in large genome cereals

Content Provider	Springer Nature Link
Author	Singh, Jaswinder Zhang, Shibo Chen, Calvin Cooper, Laurel Bregitzer, Phil Sturbaum, Anne Hayes, Patrick M. Lemaux, Peggy G.
Copyright Year	2006
Abstract	Transposable elements have certain advantages over other approaches for identifying and determining gene function in large genome cereals. Different strategies have been used to exploit the maize Activator/dissociation (Ac/Ds) transposon system for functional genomics in heterologous species. Either large numbers of independent Ds insertion lines or transposants (TNPs) are generated and screened phenotypically, or smaller numbers of TNPs are produced, Ds locations mapped and remobilized for localized gene targeting. It is imperative to characterize key features of the system in order to utilize the latter strategy, which is more feasible in large genome cereals like barley and wheat. In barley, we generated greater than 100 single-copy Ds TNPs and determined remobilization frequencies of primary, secondary, and tertiary TNPs with intact terminal inverted repeats (TIRs); frequencies ranged from 11.8 to 17.1%. In 16% of TNPs that had damaged TIRs no transposition was detected among progeny of crosses using those TNPs as parental lines. In half of the greater than 100 TNP lines, the nature of flanking sequences and status of the 11 bp TIRs and 8-bp direct repeats were determined. BLAST searches using a gene prediction program revealed that 86% of TNP flanking sequences matched either known or putative genes, indicating preferential Ds insertion into genic regions, critical in large genome species. Observed remobilization frequencies of primary, secondary, tertiary, and quaternary TNPs, coupled with the tendency for localized Ds transposition, validates a saturation mutagenesis approach using Ds to tag and characterize genes linked to Ds in large genome cereals like barley and wheat.
Starting Page	937
Ending Page	950
Page Count	14
File Format	PDF
ISSN	01674412
Journal	Plant Molecular Biology
Volume Number	62
Issue Number	6
e-ISSN	15735028
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2006-09-27
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Ac /Ds Barley Remobilization Transposon tagging Plant Pathology Biochemistry Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science Medicine Agronomy and Crop Science

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