The Trihelix Transcription Factor Family Trihelix Transcription Factor Genes Where in Arabidopsis thaliana is At3g14180 active What genotypes are present in my plants
Jurenovich, Jerome, Host has reference to this Academic Journal, PHwiki organized this Journal The Trihelix Transcription Factor Family Heather Hern in addition to ez Also called GT factors. DNA binding proteins. Bind to GT elements in promoters. Bind to promoter elements required as long as light responsiveness. Regulate plant-specific processes. Specific regulatory functions largely unknown. What is a Trihelix Transcription Factor Conserved Domain found in all trihelix proteins. Trihelix Transcription Factor Genes Gene name: At3g14180 Location: Chromosome 3 Gene size: 1736 Base Pairs Protein size: 444 amino acids Protein encoded: expressed protein similar to 6b-interacting protein 1 (found in Tobacco) UTR UTR REVERSE ORIENTATION 5 3 EXON
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Where in Arabidopsis thaliana is At3g14180 active RT-PCR 05/23/06 1.5% agarose gel 1.5 hours @ 90v 10 uL EtBr Ladder INF RT + INF RT- SIL RT + SIL RT – + Control – Control UCLA Genechip Data + Control – Control What genotypes are present in my plants Ladder Plant 1 Plant 2 Plant 3 Plant 4 Plant 5 Wildtype + Control – Control At3g14180 Genotyping 1% Agarose gel 1.5 hrs @ 105 v 10 uL EtBr T-DNA bottom T-DNA top Wildtype b in addition to Ladder Plant 6 Plant 7 Plant 8 Plant 9 Plant 10 Plant 11 Plant 12 Plant 13 Plant 14 Plant 15 Plant 16 Wildtype + Control – Control 1% Agarose gel 1.5 hrs @ 105 v 10 uL EtBr At3g14180 Genotyping Wildtype: 1 Heterozygous mutant: 4 Homozygous knockout: 11 Where did the two T-DNA b in addition to s insert Ladder LBb1 LBb1 B T 5 3 BP 1 BP 1736 830 846 Location from SALK matches my sequencing results. (SALK predicted 844 BP into the gene.) My sequencing results:
What is the impact of the T-DNA inserts No visible phenotypical differences seen in homozygous knockout plants vs. wildtype plants. What is the impact of the T-DNA inserts on seed development Wildtype heart stage Mutant heart stage Wildtype late torpedo Mutant torpedo Mutant fully developed Wildtype fully developed suspensor Trihelix Transcription Factor Gene 2 Gene name: At1g54060 Location: Chromosome 1 Gene size: 1646 Base Pairs Protein size: 384 amino acids Protein encoded: expressed protein similar to 6b-interacting protein 1 (found in Tobacco) UTR 293 BP UTR 200 BP 5 3 Exon (BP 1-1646) Forward Orientation
Where in Arabidopsis thaliana is At1g54060 active Ladder Ladder INF +RT INF -RT SIL RT + SIL RT – + Control – Control 1.5 % Agarose gel 1.5 hrs @ 105 v 10 uL EtBr RT-PCR At1g54060 Genechip Data What genotypes are present in my plants Ladder Plant 1 Plant 2 Plant 3 Plant 4 Plant 5 Plant 6 Plant 7 Plant 8 Plant 9 Plant 10 Plant 12 Plant 13 Plant 14 Plant 15 Wildtype + Control Control Genotyping At1g54060 1% Agarose gel 1.5 hrs @ 105 v 10 uL EtBr Wildtype: 6 Heterozygous T-DNA: 3 Homozygous knockout: 5 Where did the T-DNA insert LBb1 BP 635 5 3 BP 1 BP 1646 My sequencing results: My results matched the SALK institute expected insert site of 640 BP into the gene.
What is the impact of the T-DNA inserts on phenotype Plant 6 Silique 1 Plant 6, Silique 1, Top Plant 6, Silique 1, Bottom Plant 6, Silique 1, Dissected T-DNA impact continued Plant 6, Silique 2, Dissected Plant 6, Silique 1, White seeds Plant 6, Silique 2, Bottom Plant 6, Silique 2, 12.5x How does upstream regulatory activity show where the protein is translated iProof PCR At3g14180 Ladder Promoter Region + Control – Control 1% Agarose gel 1.5 hrs @ 105 v 10 uL EtBr EcoRI Digest At1g54060 Ladder Sample 1 Sample 2 Sample 3 Sample 4
Acknowledgement s Dr. Robert Goldberg Dr. Anthu Bui Mike Gavino Jonathan Russell Garima Yagnik Tomo Kawashima Jessica Luke Dr. Xingjun Wang Lab members!
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