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221.S16
Yeast GAL system (simplified)
GAL4 GAL80
translation translation
OFF
UAS
OFF
UAS
galactose + GAL3
-GAL3(galactose)
RNAP II ON
UAS
GAL2
GAL7
GAL10
221.S16 PNAS | 2002 | vol. 99 | no. 13 | 8548-8553
Yeast GAL system (simplified)
221.S16
Identification of cis-acting elements using reporter genes
proteinfusion
reporter gene
A gene whose phenotypic expression is easily monitored
• lacZ
• GFP
221.S16
Identification of cis-acting elements using reporter genes
and deletions
dsDNA
221.S16
DNase footprinting
DNase I:
a sequence nonspecific
endodeoxyribonuclease
221.S16
Biochemical evidence: DNase footprinting
bottom short
221.S16
Upstream Activating Sequence (enhancer)
of the yeast GAL1 and GAL10 genes
UDP-glucose-4-epimerase galactokinase
221.S16
High resolution genetic mapping
221.S16
Yeast GAL4 protein structure and function
DNA binding
dimerization
N-terminal 65 aa
http://gibk26.bse.kyutech.ac.jp/jouhou/image/gallery.html
https://youtu.be/WyU2v7HT6bw?t=3s
http://www.youtube.com/watch?v=GRL_rdB30GY&feature=relmfu
221.S16
Mutations that affect specific functions often
map in specific protein domains
221.S16 Hartwell et al. (2008) Fig 17.8c Krebs et al. (2008) Fig 26-16
GAL4 transcription factor with function-specific domains
1-65
221.S16
GAL4 transcription factor with function-specific domains
221.S16