transposable elements
description
Transcript of transposable elements
transposableelements
BIOL20332/20972
GENETICS / Dev. Biol. RSM
MODULE 3
Transposons as virus-like parasitic elements?
Making use of transposons
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wt stained with anti-slit
Analysing gene regulation
Analysing gene regulation
fly
molecularcloning
Analysing gene regulation
Analysing gene regulation
wt stained with anti-slit
slit-lacZ stained with anti-βGal
Analysing gene regulation
Groups larval genotype staining comments
A B C D A-lacZ 2hr 37ºC, then o.N. 26ºC 4 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, then o.N. 26ºC 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-lacZ 2hr 37ºC, then o.N. 26ºC 4 groups
How to generate transgenic fly lines with
these constructs?
Making transgenic
flies
Making transgenic
flies
Important improvements of transposable elements
that have shaped work with Drosophila
P
Enhancer trap experiments
Enhancer trap experiments
Difference: enhancer is not transgenic but endogenous
Rescue experiment of enhancer trap lines to clone the adjacent gene
The Gal4/UAS system
X-Gal4 ; UAS-lacZ
Groups larval genotype staining comments
A B C D A-lacZ 2hr 37ºC, then o.N. 26ºC 4 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, then o.N. 26ºC 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-lacZ 2hr 37ºC, then o.N. 26ºC 4 groups