TILLING and Point Mutation Detection
Maize reverse genetics is extremely powerful because of the wealth of active transposable elements still residing in the maize genome. In addition to serving as molecular tags for mutated genes, these elements tend to knock out genes into which they insert. Once an initial, reference allele has been identified, it facilitates obtaining an allelic series of mutations within a gene that have a range of phenotypes. Such an allelic series can provide great insight into the function of the gene and its cognate protein. In addition to spontaneous mutations, this allelic series is typically the result of chemical mutagenesis to create point mutations at locations throughout the gene. Forward, “targeted” mutagenesis has long been the way to do this. Mutagenizing with ethyl methane sulfonate (EMS) and then crossing onto a line homozygous for a reference allele, any mutant progeny that arise in the F1 represent new mutations in the gene of interest. While effective, a new mutagenesis is required for each gene of interest. The capacity to screen any mutagenized population using reverse genetic techniques for any gene of interest is therefore valuable as well.
KeywordsMaize Genome Ethyl Methane Sulfonate Reference Allele Allelic Series Single Base Change
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