Directed gene expression in Drosophila

1
Directed gene expression in Drosophila
(GAL4/UAS system)
2
Mosaic analysis using FLP recombinase
Producing and marking clones of cells with
different doses of mutation A
FLP
Producing and marking clones of cells with
expression of gene A -- FLP-out technique
FLP
3
FLP-out technique
constitutive promoter
gene of interest
GOI expression driven by constitutive promoter in
yellow cells only
4
from http//flybrain.uni-freiburg.de/Flybrain/ht
ml/contrib/1997/itok1997a/fig2.html
5
Chromatin immunoprecipitation (ChIP)
DNA-binding proteins are crosslinked to DNA with
formaldehyde in vivo.
Isolate the chromatin. Shear DNA along with bound
proteins into small fragments.
Bind antibodies specific to the DNA-binding
protein to isolate the complex by precipitation.
Reverse the cross-linking to release the DNA and
digest the proteins.
Use PCR to amplify specific DNA sequences to see
if they were precipitated with the antibody.
from http//www.bio.brandeis.edu/haberlab/jehsite
/chip.html 2/2/2005
6
Mosaic analysis using FLP recombinase
Producing and marking clones of cells with
different doses of mutation A
7
Producing and marking clones of cells with
expression of gene a FLP-out technique