Determining Functionality of Arabidopsis Thaliana Genes in Embryo Development
The Role of Genes AT5G48150 and AT2G04890 in Arabidopsis thaliana Seed Development
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Transcript of The Role of Genes AT5G48150 and AT2G04890 in Arabidopsis thaliana Seed Development
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The Role of Genes AT5G48150 and AT2G04890
in Arabidopsis thaliana Seed Development
Jennifer HuynhJune 8, 2006
Honors Collegium 70ALProfessor Bob Goldberg
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Overview
GRAS Family
Knockout of Gene
Future Research
mRNA Accumulation
Promoter Regions
AT5G84150AT2G04890
Conclusions
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What has been already discovered about the genes?
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The GRAS Family• Name derives from the three initially
identified members– GAI, RGA, and SCR
• Functions range from meristem maintenance to regulation of hormone and light signal transduction
• PAT1 branch plays role in far red light signaling
• Homologs found in higher plants– Tomato, petunia, lily, rice, barley
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The GRAS Family Tree
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The GRAS Family• Name derives from the three initially
identified members– GAI, RGA, and SCR
• Functions range from meristem maintenance to regulation of hormone and light signal transduction
• PAT1 branch plays role in far red light signaling
• Homologs found in higher plants– Tomato, petunia, lily, rice, barley
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Gene One: AT5G48150• Scarecrow-like transcription factor 21 (SCL21)
Gene Size: 2355 bp Protein Size: 491 aa
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Gene Two: AT2G04890• Phytochrome A signal transduction 1 (PAT1)
Gene Size: 2601 bp Protein Size: 414 aa
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Where is the gene transcribed into mRNA?
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mRNA Accumulation: AT5G48150RT-PCR Results
GeneChip DataControl cDNA: 603 bp
Control Genomic: 922 bp
AT5G48150 Gene: 323 bp
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mRNA Accumulation: AT2G04890RT-PCR Results
GeneChip DataControl cDNA: 603 bp
Control Genomic: 922 bp
AT2G04890 Gene: 499 bp
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Is the upstream region of the gene responsible for the
transcription of the gene?
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Restriction-Digested Plasmid DNAAT5G48150
AT2G04890
Plasmid: 3500 bpPromoter: 3022 bpGel Bands: 1800 bp, 1200bp
Plasmid: ~3500 bp Promoter: 2923 bp
Gel Bands: 1600 bp, 1300 bp
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What happens if the genes are knocked out?
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Genotyping Insertion LinesAT5G48150
AT2G04890
Wildtype: 1796 bpSalk band: 1740 bp Found band: 1739 bp
Wildtype: 1508 bp
Salk band: 756 bp
Found band: 880 bp
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Genotyping Insertion Lines
AT5G48150
AT2G04890
Wildtype: 1796 bpSalk band: 1740 bp Found band: 1739 bp
Wildtype: 1508 bp
Salk band: 756 bp
Found band: 880 bp
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AT5G48150: SALK_064220• Wildtype: 1796 base pairs• Salk band: 1740 base pairs • Found band: 1739 base pairs
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AT2G04890: SALK_146085
• Wildtype: 1508 base pairs• Salk band: 756 base pairs• Found band: 880 base pairs
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Nomarski Photographs
Wildtype
Mutant
Globular
Globular
Heart
Heart
Torpedo
Torpedo Late Torpedo
Late Torpedo
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Conclusions• No mutant phenotype on flowers, leaves,
or siliques for either AT5G48150 or AT2G04890
• Possibilities– Genes not vital for seed development– Similar genes exist that compensate– Mutant phenotype not visible in experiments
• More research is necessary
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Future Research
• Perform RT-PCR on homozygous mutants seeds to observe transcription
• Fuse GFP marker with promoter to observe expression
• Cross homozygous mutant plants of different knockouts
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Acknowledgements
• Bob Goldberg
• Anhthu Bui
• Mike Gavino
• Jonathan Russell
• Ria Yagnik
• Brandon Le
• Xingjun Wang
• Tomo Kawashima
• Jessica Luke
• HC 70AL Classmates