HowdowefindaknockoutforAT4G37790andwhatisthisgene.pptVIP

How do we find a knockout for AT4G37790 and what is this gene’s function in seed development? What do we know about AT4G37790? Common Name: HAT-22 Homeobox leucine-zipper protein Codes for nuclear protein sized at 278 AA Regulatory DNA binding Important for transcription factor function Contains 1 homeobox domain Located on chromosome 4 of Arabidopsis Forward strand Where is HAT-22 active in Arabidopsis Thaliana and in SRB? In SRB, the gene was found to be the most active in Sample 1 (Genomic Leaf +RT), Sample 3 (Genomic Stem +RT), and Sample A (Isolated Leaf +RT). In Arabidopsis, the gene is most active in lec1 Stem (signal detection 4087.3 P), the WT Stem (3170 P), lec1 Leaves (2678.5 P), and WT Leaves (2491 P). What is the structure of AT4G37790? Which superpool from the Madison Facility contains a t-DNA insert in AT4G37790? Superpool 3 contained the correct hit, as shown in the FW autoradiogram above. The RV autoradiogram contained no hits. From the gel, the target segment was sized at ~800 bp. Which DNA pool contains a t-DNA insert in AT4G37790? Amplify PCR products from 9 DNA pools belonging to Superpool 3. Use FW JL202 primers and a + control, Superpool 3. Can we find a PCR product with the same size as the hit segment from our superpool screening (~800 bp)? The primers amplify an ~800 bp segment in DNA Pool 4. How do SALK Lines help us see how AT4G37790 functions in seed development? SALK Line Used: 085964.56.00 T-DNA Insertion at -312 bp Complementary Orientation What are the genotypes of the SALK plants with respect to the t-DNA insert? Where do we go from here? SALK LINES: Collect seeds from heterozygous mutant plants and attempt to raise homozygous mutants. If homozygous mutants are obtained, knockout is not embryo lethal. However, we can PHENOTYPE the plants to see whether abnormalities give us clues to the importance of AT4G37790 in seed development. Salik examination MADISON FACILITY: Continue to search for