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doi:10.1534/genetics.104.033738
A more recent version of this article appeared on February 1, 2005.
REGULAR RESEARCH PAPERS |
Distribution of Activator (Ac) Throughout the Maize Genome for Use in Regional Mutagenesis
Judith M. Kolkman 1, Liza J. Conrad 2, Phyllis R. Farmer 1, Kristine Hardeman 3, Kevin R. Ahern 1, Paul E. Lewis 1, Ruairidh J. H. Sawers 1, Sara Lebejko 3, Paul Chomet 3 and Thomas P. Brutnell 1*
1 Boyce Thompson Institute
2 Cornell University
3 Monsanto Company
* To whom correspondence should be addressed. E-mail: tpb8{at}cornell.edu.
Submitted on July 19, 2004
Revised on September 17, 2004
Accepted on 28 October 2004
A collection of Activator (Ac) containing, near-isogenic W22 inbred lines has been generated for use in regional mutagenesis experiments. Each line is homozygous for a single, precisely positioned Ac element and the Ds reporter, r1-sc:m3. Through classical and molecular genetic techniques, 158 transposed Ac elements (tr-Acs) were distributed throughout the maize genome and 41 were precisely placed on the linkage map utilizing multiple recombinant inbred populations. Several PCR techniques were utilized to amplify DNA fragments flanking tr-Ac insertions up to 8 kb in length. Sequencing and database searches of flanking DNA revealed the majority of insertions are in hypomethylated, low or single copy sequences indicating an insertion site preference for genic sequences in the genome. However, a number of Ac transposition events were to highly repetitive sequences in the genome. We present evidence that suggests Ac expression is regulated by genomic context resulting in subtle variations in Ac-mediated excision patterns. These tr-Ac lines can be utilized to isolate genes with unknown function, to conduct fine-scale genetic mapping experiments and to generate novel allelic diversity in applied breeding programs.
Key Words: Ac, maize, mutagenesis, transposition, transposon tagging
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