Transcript The GRAS Transcription Factor Family

The Role of Genes
AT5G48150 and AT2G04890
in Arabidopsis thaliana
Seed Development
Jennifer Huynh
June 8, 2006
Honors Collegium 70AL
Professor Bob Goldberg
Overview
AT5G84150
AT2G04890
GRAS
Family
mRNA
Accumulation
Future
Research
Promoter
Regions
Conclusions
Knockout
of Gene
What has been already
discovered about the genes?
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
The GRAS Family Tree
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
Gene One: AT5G48150
•
Scarecrow-like transcription factor 21 (SCL21)
Gene Size: 2355 bp Protein Size: 491 aa
Gene Two: AT2G04890
•
Phytochrome A signal transduction 1 (PAT1)
Gene Size: 2601 bp Protein Size: 414 aa
Where is the gene
transcribed into mRNA?
mRNA Accumulation: AT5G48150
RT-PCR Results
Control cDNA: 603 bp
GeneChip Data
Control Genomic: 922 bp
AT5G48150 Gene: 323 bp
mRNA Accumulation: AT2G04890
RT-PCR Results
Control cDNA: 603 bp
GeneChip Data
Control Genomic: 922 bp
AT2G04890 Gene: 499 bp
Is the upstream region of the
gene responsible for the
transcription of the gene?
Restriction-Digested Plasmid DNA
AT5G48150
Plasmid: ~3500 bp
Promoter: 2923 bp
Gel Bands: 1600 bp, 1300 bp
Plasmid: 3500 bp
Promoter: 3022 bp
Gel Bands: 1800 bp, 1200bp
AT2G04890
What happens if the genes
are knocked out?
Genotyping Insertion Lines
AT5G48150
Wildtype: 1508 bp
Salk band: 756 bp
Found band: 880 bp
Wildtype: 1796 bp
Salk band: 1740 bp
Found band: 1739 bp
AT2G04890
Genotyping Insertion Lines
AT5G48150
Wildtype: 1508 bp
Salk band: 756 bp
Found band: 880 bp
Wildtype: 1796 bp
Salk band: 1740 bp
Found band: 1739 bp
AT2G04890
AT5G48150: SALK_064220
• Wildtype: 1796 base pairs
• Salk band: 1740 base pairs
• Found band: 1739 base pairs
AT2G04890: SALK_146085
• Wildtype: 1508 base pairs
• Salk band: 756 base pairs
• Found band: 880 base pairs
Nomarski Photographs
Globular
Heart
Torpedo
Late Torpedo
Wildtype
Globular
Heart
Mutant
Torpedo
Late Torpedo
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
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
Acknowledgements
•
•
•
•
•
Bob Goldberg
Anhthu Bui
Mike Gavino
Jonathan Russell
Ria Yagnik
•
•
•
•
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Brandon Le
Xingjun Wang
Tomo Kawashima
Jessica Luke
HC 70AL Classmates