transposable elements
21
transposable elements BIOL20332/20972 GENETICS / Dev. Biol. RSM MODULE 3 The University of Manchester Faculty of Life Sciences Andreas Prokop
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BIOL20332/20972 GENETICS / Dev. Biol. RSM MODULE 3. The University of Manchester. transposable elements. Faculty of Life Sciences. Andreas Prokop. Making use of transposons !!!!. Transposons as virus-like parasitic elements?. A. Prokop. Analysing gene regulation. - PowerPoint PPT Presentation
Transcript of transposable elements
transposableelements
BIOL20332/20972
GENETICS / Dev. Biol. RSM
MODULE 3
The
Uni
vers
ityof
Man
ches
ter
Fac
ulty
of
Life
Sci
ence
s
Andreas Prokop
Transposons as virus-like parasitic elements?
Making use of transposons
!!!!
A.
Pro
kop
wt stained with anti-slit
Analysing gene regulation
A.
Pro
kop
Analysing gene regulation
fly
molecularcloning
A.
Pro
kop
Analysing gene regulation
A.
Pro
kop
Analysing gene regulation
A.
Pro
kop
wt stained with anti-slit
slit-lacZ stained with anti-βGal
Analysing gene regulation
A.
Pro
kop
Groups larval genotype staining comments
A B C D A-lacZ2hr 37ºC, then
o.N. 26ºC4 groups
E F G H B-lacZ 2hr 37ºC 4 groups
I J K L C-lacZ 2hr 26ºC 4 groups
M N O D-lacZ2hr 37ºC, then
o.N. 26ºC3 groups
P Q R E-Gal4;UAS-lacZ 2hr 37ºC 3 groups, stock a bit weak
S T U F-Gal4;UAS-lacZ 2hr 37ºC 3 groups
V W X Y G-lacZ2hr 37ºC, then
o.N. 26ºC4 groups
A.
Pro
kop
How to generate transgenic fly lines with
these constructs?
A.
Pro
kop
Making transgenic
flies
A.
Pro
kop
Making transgenic
flies
A.
Pro
kop
Important improvements of transposable elements
that have shaped work with Drosophila
A.
Pro
kop
P
Enhancer trap experiments
A.
Pro
kop
Enhancer trap experiments
Difference: enhancer is not transgenic but endogenous
A.
Pro
kop
Rescue experiment of enhancer trap lines to clone the adjacent gene
A.
Pro
kop
The Gal4/UAS system
A.
Pro
kop
X-Gal4 ; UAS-lacZ
A.
Pro
kop
Groups larval genotype staining comments
A B C D A-lacZ2hr 37ºC, then
o.N. 26ºC4 groups
E F G H B-lacZ 2hr 37ºC 4 groups
I J K L C-lacZ 2hr 26ºC 4 groups
M N O D-lacZ 2hr 37ºC 3 groups
P Q R E-Gal4;UAS-lacZ 2hr 37ºC 3 groups, stock a bit weak
S T U F-Gal4;UAS-lacZ 2hr 37ºC 3 groups
V W X Y G-lacZ2hr 37ºC, then
o.N. 26ºC4 groups
A.
Pro
kop
You performed an enhancer trap screen: what to do next?
A.
Pro
kop
and identify the gene
use the line as marker for
experiments
perform gain-of-function
experiments
A.
Pro
kop
Carry out loss-of-function studies
A.
Pro
kop
