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