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Sequence the DNA of 7000-9000 unique A. flavus Expressed Sequence Tags (EST's) and perform data analysis and annotation of the DNA sequence for preparation of microarrays to be used in generating data necessary for incorporating genetic resistance to aflatoxin contamination into crops.
Determination of A. flavus Expressed Sequence Tags (EST's)can help to identify and measure expression of the majority (7000-9000 genes) of the genes expressed in the fungal genome. Fragments of these genes will be amplified for fabricating microarrays for quantifying their expression. A. flavus EST and genomic databases will provide more specific information on which critical regulatory components and genes involved in aflatoxin biosynthesis and/or fungal growth should be targeted through selection of expressed genes in the plant encoding inhibitors of fungal growth and/or aflatoxin formation. To obtain 90% of expressed genes in A. flavus, 5 X sequence coverage would be necessary. However, on a practical basis, this goal will be achieved by 2-3 X coverage with a normalized library made commercially in order to further reduce the cost. The annotated sequence information will be available to the public through deposition into the GenBank Data base. In the meantime, EST information will be made available for ARS research and possibly for sharing with outside research groups.