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Identify dys fusion cell enhancer. Fusion cell gene profiling. dys-gal4::UAS-CD8-GFP. btl-gal4::UAS-CD8-GFP. Identify dys target gene. dys-gal4::UAS-CD8-GFP. dys-gal4::UAS-CD8-GFP ; dys/dys. Molecular mechanism of target gene regulation by Dys::Tgo. Dys::Tgo activation of CG13196.
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Identify dys fusion cell enhancer • Fusion cell gene profiling dys-gal4::UAS-CD8-GFP btl-gal4::UAS-CD8-GFP • Identify dys target gene dys-gal4::UAS-CD8-GFP dys-gal4::UAS-CD8-GFP ; dys/dys • Molecular mechanism of target gene regulation by Dys::Tgo • Dys::Tgo activation of CG13196 • Dys:Tgo activation of CG15252 • Identify mbo and CG15252 enhancer
dys genomic structure FTF Q72L FTF T31N Exon1 2 3 4 5 6 7 8 9 10 LP05454 3.8kb F2F elution 1.7kb 2.7kb 3.8kb 3.8kb
Expression patterns of CG13196 enhancer and TCGTG enhancer 4xTCGTG GFP Dys CG13196 GFP Dys Early S15 S16
Ectopic expression of dys can activate CG13196 and TCGTG enhancer ectopically CG13196 Dys 4xTCGTG GFP Dys S15 S16
CG15252 5’ upstream TCGTG TCGTG TCGTG Exon1 3.2kb 0.8 kb TCGTG cluster 9.0 kB 5’ upstream
Spalt is not required for CG15252 activation by dys in DT CG15252 Dys CG15252 Dys btl-gal4;UAS-dys btl-gal4;UAS-spalt;UAS-dys Need to do: CG15252 expression in Spalt mutants
a-Dys mbo-lacZ btl-gal4, UAS-dys Ectopic dys can activate mbo-lacZ ectopically
mbo 3’ TCGTG ACGTG 407bp
Conclusion • dys target genes can only be turn on at late stages (late 14 or early 15) • There is no transcriptional inhibition at early stage • There are other transcriptional activators turns on at late stages or changes in chromatin structure. • Spalt is not required for CG15252 expression