At5g A mutant phenotype? Emily Eder HC70AL - Spring 2005

Arabidopsis Gene At5g65640 Plant genotyping Sequencing and T-DNA insert First observation of mutant phenotypes Nomarski RNA extraction – microarray – expression

Genotyping 4 Homozygous, 0 Heterozygous, and 8 Wild Type Plants Homozygous

Sequencing the Mutant Band Why is it necessary to sequence the band? We must verify the T-DNA insertion site

AT5g Base Pairs in Length Codes for basic helix-loop-helix (bHLH) protein 146

Mutant Phenotypes

Mutant Phenotypes cont.

Using Microscope to Examine Phenotypes

Microscope Examination cont.

Mutant Nomarski

Wild Type Nomarski

Using Gel Electrophoresis to Determine RNA Expression

Gel Electrophoresis to Determine RNA Expression

Using a Genechip (Microarray) to Determine RNA Expression

Arabidopsis Gene AT5g Homozygous Mutant Plants - all with visible phenotype 1 T-DNA insert - forward orientation RNA expression revealed a large amount of RNA present in the seed

What’s next? Can we be sure that this is an actual mutant phenotype? More research is necessary