|Tan, Bao Cai|
|Porch, Timothy - Tim|
Submitted to: Plant and Animal Genome Conference
Publication Type: Abstract Only
Publication Acceptance Date: 10/10/2003
Publication Date: 1/20/2004
Citation: Settles, M., Hannah, C.L., Kock, K.E., Baier, J., Latshaw, S., Avigne, W., Tan Bao, C., Suzuki, M., Porch. T.G., Becraft, P., Larkins, B., Messing, J., McCarty, D.R., 2004. Functional Genomics of endosperm development in maize (abstract). Plant and Animal Genome Conference.
Technical Abstract: The Maize Endosperm Functional Genomics Project is developing tools for the molecular cloning of seed mutants in maize. Specifically, we are developing normalized and subtracted ESTs from endopserm cDNA libraries, identifying transposon-tagged endosperm mutants from the mutagenic inbred, UniformMu, and cloning and sequencing the transposon insertion sites from a subset of these seed mutants. These transposon insertions represent both random gene disruptions and insertions in genes that cause endosperm phenotypes. So far, we have achieved or exceeded the numerical goals for these key deliverables: 1) 23,348 subtraction normalized endosperm EST’s have been deposited in Genbank, 2) 2,025 primary mutant isolates have been generated with the UniformMu transposon population. Ongoing efforts are directed at adding value to these resources by 1) completing full length cDNA sequences for a non-redundant set of endosperm cDNA’s, 2) confirming heritability of all mutants with the goal of establishing at least 2,200 independent confirmed mutants. With the benefit of a supplemental award for MuTAIL sequencing, we have shifted from a hybridization based strategy to a sequence based strategy for molecular analysis of the mutants. Toward that goal, we completed construction of MuTAIL clone libraries from 90 mutants and we are sequencing of 344 sub-libraries from these mutants. To date, 25,509 MuTAIL sequences have been deposited in the GSS database. Major bioinformatics efforts include construction of a centralized data management system for the Florida project, development of a suite of specialized database tools for mutant scoring, imaging and pedigree data, implementation of a barcode sample tracking system, and construction of an annotation database for MuTAIL sequences that will facilitate identification of sequence tagged genes and candidates for endosperm mutants. Several mutant genes have been successfully cloned by Project collaborators using MuTAIL (vp13, vp15, opaque16, and wpk) illustrating the power of this PCR strategy for cloning Mu tagged genes.