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Arabidopsis thaliana gene At3g50060

Arabidopsis thaliana gene At3g50060. A Gene Story HC70AL Spring 2004 Gloria Chu. Gene Number: At3g50060 Gene Name: AtMYB77 or AtMYBr2 Function: myb transcription factor. The Gene. 5’ 1. 3’ 1135. UTR. Gene AT3G50060. Exon. Scarlet Runner Bean EST.

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Arabidopsis thaliana gene At3g50060

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  1. Arabidopsis thaliana gene At3g50060 A Gene Story HC70AL Spring 2004 Gloria Chu

  2. Gene Number: At3g50060 Gene Name: AtMYB77 or AtMYBr2 Function: myb transcription factor The Gene 5’ 1 3’ 1135 UTR Gene AT3G50060 Exon Scarlet Runner Bean EST The Protein • 508 bps • Aligns with At3g50060 for approximately 216 bps • - from +152 to +367 of gene • From suspensor of SRB • PCSC11470: similar to At4g37260 (AtMYB73) • Codes for 301 Amino Acids • Similar protein found in Solanum tuberosum “The hallmark of these proteins is a highly conserved DNA-binding domain composed of tandem repeats of a helix-turn-helix motif (light blue) that fit into the major groove of the DNA double helix (red and yellow).”* * http://www.stanford.edu/group/lipsick/whatsmyb%20short.htm

  3. Gene Activity Odd Lanes are +RT Even Lanes are -RT controls Control Band

  4. The Big Question Is my gene vital for seed development in Arabidopsis plants? The Big Question Reworded for the Sake of Experimentation Does knocking out my gene cause an embryo lethal mutation?

  5. Madison Screen Results Round 1: Superpools Round 2: DNA Pools • DNA Pools from Superpool 22 • 2nd DNA Pool (Pool 191) shows clear band at approximately correct size of At3g50060 fragment (1496 bps) Superpools amplified using gene specific FW primer Superpool 22 Superpool 2 L 1 2 3 4 5 6 7 8 9 C L Superpools amplified using gene specific RV primer JL202 Primer 22 Fw Gene-specific Fw primer

  6. SALK InsertIons HITS SALK_067655.56.00.x TYPE SALK T-DNA [TDNA] EVAL e-139 LOCN Exon COOR C/18569060-18569313 HITS SALK_055373.34.15.x TYPE SALK T-DNA [TDNA] EVAL 5e-88 LOCN 300-UTR5 COOR C/18569907-18570052

  7. Graphical Depiction of Gene At3g50060 and T-DNA Insertions 055373 067655 22 FW 2FW 5’ 3’ FW RV Primers Exon Intron SALK Madison Locations of T-DNA Insertions Madison SALK Superpool 2Fw: +1117 (3’UTR) Line #067655: +776 (exon) Superpool 22Fw: +965 (3’UTR) Line #055373: +37 (5’ UTR)

  8. SALK Plant Genotypes L 1 2 3 4 5 6 7 8 9 WT Neg. Control Gel 1: Wild-Type Allele Gene-specific Fw and Rv primers Gel 2: Mutant Allele Gene-specific Rv primer and LBb1 L 1 2 3 4 5 6 7 8 9 WT N P

  9. SALK Plant Phenotypes Wild Type Plant (homozygous WT) COMING SOON…Due to technical difficulties Mutant (Knockout) Plant (homozygous mutant) COMING SOON…Due to technical difficulties

  10. Back to the Big Questionand Where To Go From Here Knocking out gene At3g50060 does not cause an embryo lethal mutation Photo: A Greenhouse; not the UCLA greenhouse; just a greenhouse

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