AT5g67300

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AT5g67300

• Emily Eder
• HC70AL - Spring 2005

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Arabidopsis Gene AT5g67300

• Plant genotyping
• Sequencing and T-DNA insert
• RNA extraction and expression
• Using gel electrophoresis
• Using a genechip (microarray)
• Promoter Region

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Genotypes

• All Homozygous Mutants

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Genotypes cont.
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AT5g67300
1216 Base Pairs in Length, Forward Orientation
Codes for MYB Family Transcription Factor
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Using Gel Electrophoresis to Determine RNA
Expression
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Using a Genechip (Microarray) to Determine RNA
Expression
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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

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Promoter Sequence via Finch TV
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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

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Using SP6 Primer

• No mutations

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Using T7 Primer

• 1 mutation

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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