Arabidopsis Thaliana A Study of Genes and Embryo Development By Garen Polatoglu

A T 1 G Gene Gene Length: 1511 bp Translated Region: 786 bp PCR Product: 1004 bp Encoded Protein Size: 261 amino acids Putative AP2 domain containing transcription factor

Genotyping Homozygous Mutants (~1.2kb) –Plants 1, 2, 4, 6, 7 Wild Type (~1kb) –Plants 3, 5, 8

tDNA Insertion Site AT1G22190 Excised Band Sequence Alignment at 477 bp Does not agree with proposed SALK_139727

Gene Expression RT-PCR –Results indicate that the gene is active in both Leaf and Silique. –Bands in –RT lanes and Negative Control erroneous

Gene Expression AT1G22190 Genechip Data From the laboratory of Dr. Goldberg

Promoter Region Promoter Region is immediately upstream of AT1G22190 Size: 3002 bp Restriction Sites: EcoRI –GAATTC –Promoter cut into 1950 bp bp

Promoter Analysis The promoter clone of Colony 1 was sequenced for verification purposes The sequence did not match exactly with that of TAIR’s Entire promoter (~3kb) was not sequenced EcoRI Promoter Digestion Gel 3.5kb 2.0kb 1.0kb Plasmid

Plant Phenotypes No variations were observed between the homozygous mutant plants and wild type plants Seeds of both genotypes were observed under a Nomarski microscope –No differences were noted

Nomarski Images Wild Type SeedMutant Seed

What’s next? Causes for no phenotypic effect of knockout gene could be varied –Genetic Redundancy –Phenotype more apparent in other conditions Multiple knockouts of similar genes may induce a varied phenotype