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Arabidopsis Thaliana

Arabidopsis Thaliana. A Study of Genes and Embryo Development By Garen Polatoglu. A T 1 G 2 2 1 9 0. Encoded Protein Size: 261 amino acids Putative AP2 domain containing transcription factor. Gene Gene Length: 1511 bp Translated Region: 786 bp PCR Product: 1004 bp.

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Arabidopsis Thaliana

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

  2. A T 1 G 2 2 1 9 0 • Encoded Protein • Size: 261 amino acids • Putative AP2 domain containing transcription factor • Gene • Gene Length: 1511 bp • Translated Region: 786 bp • PCR Product: 1004 bp

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

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

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

  6. Gene ExpressionAT1G22190 Genechip Data From the laboratory of Dr. Goldberg

  7. Promoter Region • Promoter Region is immediately upstream of AT1G22190 • Size: 3002 bp • Restriction Sites: EcoRI • GAATTC • Promoter cut into 1950 bp + 1052 bp

  8. 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 2.0kb 1.0kb 3.5kb Plasmid

  9. 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

  10. Nomarski Images Wild Type Seed Mutant Seed

  11. 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

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