1. Emily Eder HC70AL - Spring 2005
AT5g67300
Emily Eder
HC70AL - Spring 2005

2. Arabidopsis Gene AT5g67300 Plant genotyping
Sequencing and T-DNA insert
RNA extraction and expression
Using gel electrophoresis
Using a genechip (microarray)
Promoter Region

3. All Homozygous Mutants
Genotypes
All Homozygous Mutants

4. Genotypes cont.

5. AT5g67300 1216 Base Pairs in Length, Forward Orientation
Codes for MYB Family Transcription Factor

6. Using Gel Electrophoresis to Determine RNA Expression

7. Using a Genechip (Microarray) to Determine RNA Expression

8. Promoter Region
Why do we need to clone the promoter region of our gene?
More accurate gene expression
Why do we need to verify the sequence of the promoter region?
Ensure that DNA sequence was not mutated

9. Promoter Sequence via Finch TV

10. Alignment Blast How can we verify that our sequence was not mutated?
By blasting the two sequences together
Verify mutations for both SP6 and T7

11. Using SP6 Primer
No mutations

12. Using T7 Primer
1 mutation

13. Arabidopsis Gene AT6g67300 All Homozygous Mutant Plants
One T-DNA insert with forward orientation
RNA expression much higher in the leaf than the seed/silique
Promoter region successfully cloned