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Do AT4G13640 and AT3G24120 Play a Significant Role in Seed Development for Arabidopsis Thaliana ?

Do AT4G13640 and AT3G24120 Play a Significant Role in Seed Development for Arabidopsis Thaliana ?. By Jordan Fischer June 8, 2006 Professor Goldberg hc70al. Transcription factors are proteins involved in the regulation of gene expression.

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Do AT4G13640 and AT3G24120 Play a Significant Role in Seed Development for Arabidopsis Thaliana ?

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  1. Do AT4G13640 and AT3G24120 Play a Significant Role in Seed Development for Arabidopsis Thaliana? By Jordan Fischer June 8, 2006 Professor Goldberg hc70al

  2. Transcription factors are proteins involved in the regulation of gene expression. Bind to the promoter region upstream of gene and either facilitate or inhibit transcription. Control and regulate gene expression. Activated by: Physiological stimuli. Therapeutically. Pathological stimuli. What Is a Transcription Factor? Examples of Transcription Factors

  3. Recently categorized GARP super family of transcription factors defined by G2 in maize; the Arabidopsis RESPONSE REGULATOR-B proteins; and the PHOSPHATE STARVATION RESPONSE1 protein of Chlamydomonas. G2 function is specifically committed to the differentiation of bundle sheath cell chloroplasts in C4 leaf blades. Act as transcriptional regulators of cell-type differentiation processes. Likely that GLK proteins act as transcriptional regulators of chloroplast development. What Is the Significance of theG2-like Transcription Factor Family?

  4. What is the Structure of Gene AT4G13640?

  5. What is the Structure of Gene AT3G24120?

  6. AT4G13640 AT3G24120 Protein Type MYB MYB Size (bp) 2039 2436 Size (aa) 292 295 # Exons 6 6 # Introns 5 5 Most Active 24-Hr. Seed Ovule and 24-Hr. Seed What Other Information is Important Pertaining to These Genes?

  7. According to Gene Chip, Where are These Genes Active in the Arabidopsis Plant? Gene AT3G24120 Gene AT4G13640 Control (Gene AT1G74840)

  8. Are the Results from RT-PCR Consistent with Those of Gene Chip? AT3G24120 AT4G13640 Yes, according to Gene Chip and RT-PCR both genes are active in the leaf and silique of the Arabidopsis plant. Leaf +RT Leaf -RT Silique +RT Silique -RT (+) Control Leaf +RT Leaf -RT Silique +RT Silique -RT (+) Control

  9. Did My Experiments Agree With Salk’s Regarding the T-DNA Insert Location?

  10. What Genotyping Data Was Collected For Gene AT4G13640? 1st Batch 2nd Batch 1 Kb Ladder Control (-) Control (+) Plant #18 Plant #17 Plant #16 Plant #15 Plant #14 Plant #13 Plant #12 Plant #11 Plant #10 Plant #9 Plant #8 Plant # 7 1 Kb Ladder Control (-) Control (+) Plant #6 Plant #5 Plant #4 Plant #3 Plant #2 Plant #1 1 Kb Ladder Wild Type Band Mutant Type Band

  11. What Genotyping Data Was Collected For Gene AT3G24120? RV & LBb1 FW & LBb1 FW & RV LBb1 Cont. Control (-) Control (+) Plant #17 Plant #16 Plant #15 Plant #14 Plant #13 Plant #12 Plant #11 1 Kb Ladder Control (-) Control (+) Plant #17 Plant #16 Plant #15 Plant #14 Plant #13 Plant #12 Plant #11 1 Kb Ladder LBb1 Control Control (-) Control (+) Plant #17 Plant #16 Plant #15 Plant #14 Plant #13 Plant #12 Plant #11 Wild Type Band Mutant Type Band

  12. AT4G13640 AT3G24120 Wild/Wild Mutant/Mutant Heterozygous TOTAL What is the Summary of Genotyping Results for Genes AT4G13640 and AT3G24120? 5 8 5 2 5 0 7 18

  13. How Did the Phenotypes of Homozygous Mutant and Wild Type Plants Compare? Mutant Wild Type

  14. How is Promoter Cloning Used in this Experiment? AT4G13640 Colony # 4 Colony #3 Colony #1 1 Kb Ladder (-) Control (+) Control Amp. Region 1 Kb Ladder AT3G24120 (-) Control (+) Control Amp. Region 1 Kb Ladder Colony # 4 Colony #3 Colony #2 Colony #1 1 Kb Ladder

  15. What Conclusions Can Be Made Based on these Experiments? • Genes AT4G13640 and AT3G24120 are not active in seed development to the extent where a knock out of either gene will be lethal to seed development. • A knockout of these genes causes no observable phenotypical differences compared to the wild type.

  16. What Further Experiments Should be Done on Genes AT4G13640 and AT3G24120? • Use GFP as a marker to help to determine where in the plant the promoter regions activate transcription. • Cross homozygous mutant plants from both knock out lines and determine if a complete knock out of both genes is fatal to seed development or if this creates any phenotypic differences. • Use Nomarksi microscope to check for phenotypical differences in seed development.

  17. Acknowledgements Professor Golberg Jessica Luke Anhthu Bui Brandon Le Xingjun Wang Tomo Kawashima Jonathan Russell Ria Yagnik Mike Gavino

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