Are At1g08810 and At3g50060 Important to Arabidopsis Seed Development? Jonathan Russell June 9, 2005 HC70AL

What are the Structures of My Genes? At1g08810 At3g50060

What Information is Important? At1g08810 At3g50060 Protein Type MYB 60 MYB 77 Size (bp) 1,331 1,135 Size (aa) 280 301 # of Exons 3 1 # of Introns 2 Most Active Mature Green Stage Seed Post-Mature Green Stage Seed

When/Where are My Genes Active? At1g08810 At3g50060

What Activity Did Experiments Show? At1g08810 Expected Size of RT-PCR Product At1g08810 295 bp At3g50060 576 bp At3g50060

What Were My 1st Line’s Genotypes? Expected Size of PCR Product At1g08810 Wild Type 1,131 bp T-DNA 663 bp

What Were My 1st Line’s Genotypes? Expected Size of PCR Product At1g08810 Wild Type 1,131 bp T-DNA 663 bp

What Were My 2nd Line’s Genotypes? Expected Size of PCR Product At3g50060 Wild Type 1,496 bp T-DNA 1,258 bp

Genotype Summary At1g08810 At3g50060 WT / WT 16 WT/ Mut 3 Mut / Mut 2 WT/ Mut 3 Mut / Mut 2 22

Did the Gene Maps Need Revision? At1g08810: YES At3g50060: YES

Do the Mutants Show a Phenotype? Wild Type At1g08810: NO At3g50060: NO

Does Nomarski Reveal a Phenotype? Wild Type At3g50060: NO At1g08810: NO

How is Promoter Cloning Useful? Amplification of Promoter Region (Gene 1) EcoR1 Digestion of Plasmid Vector (Gene 2) Sequencing of Promoter Region At1g08810 At3g50060 SP6 T7 Matches 629 619 463 785 Ignored 8 86 Mismatch 1 4 Expected Size of Promoter Region At1g08810 1,480 bp At3g50060 3,153 bp

What Conclusions Can be Drawn? My experiments did not reveal a mutant phenotype for either At1g08810 or At3g50060 Possible Explanations… These genes are not vital for development These genes are vital for development, but similar genes exist that can take over their functions A mutant phenotype exists, but was not visible in my specific experiments