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[Drosophila] Help: Gal4 and expression of genes with many/long introns

Claire Thomas via dros%40net.bio.net (by gxt5 from psu.edu)
Tue Sep 12 15:26:57 EST 2017


Kevin,

Gal4 - alone effects are well documented (but mostly ignored in my 
experience) e.g.:

Genet. Mol. Res. 2 (1): 43-47 (2003)

Fly (Austin). 2008 Mar–Apr; 2(2): 92–98.

Claire



On 9/12/17 1:53 PM, Sandra Schulze wrote:
> Hi Kevin,
>
> If  I understand you correctly, you are wondering if the expression of GAL4
> alone has the effect of upregulating certain genes? As if, in the promoter
> regions of these genes, there is a sequence recognized by some kind of
> transcription factor that GAL4 might resemble?
>
> I have not had quite the same experience, but I can tell you that I am in
> the middle of a survival experiment (testing for life-span extension),
> where in my pilot experiment, driving with GAL4 appeared to extend life
> span. In other words, my negative control that involved UAS GAL4 extended
> lifespan as long as my experimental.
>
> I should know in 200 short days whether this can be repeated. There were
> lots of problems with the pilot regarding genetic background, which have
> been corrected in the current experiment.
>
> I have often wondered, though, how the expression of GAL4 affects other
> genes. It is a long-utilized and hallowed tool, and negative controls
> abound, but I still wonder.
>
> Are you going to try and see if this effect can be reproduced with other
> drivers?  In isogenic backgrounds?
>
> Cheers,
>
> S
>
> On Tue, Sep 12, 2017 at 9:03 AM, Kevin Van Der Graaf <GraafK from cardiff.ac.uk>
> wrote:
>
>> Hi all,
>>
>> I have some interesting data that we don't understand, and we are
>> interested in hearing if anyone else has seen something similar.
>>
>> We have a mutant where spliced genes with many and long introns are
>> down-regulated compared to WT. We used the UAS/Gal4 system to rescue our
>> mutant.
>> However, in the control reactions we noticed that Gal4 alone might have an
>> effect on the expression of these genes.
>>
>> To give you an example, we compared a wild-type with armgal4;Sb/Tm3
>> genotype and saw that these many/long intron genes were highly up-regulated
>> in the armgal4;Sb/Tm3. When this is in the mutant background the expression
>> of the long intron genes is rescued, even with no UAS-construct present.
>>
>> Has anyone else seen this affect where just gal4 affects the expression of
>> genes (in particular genes with many/long introns)?
>>
>> Many thanks,
>>
>> Kevin van der Graaf
>> PhD student
>> Cardiff University
>> graafk from cf.ac.uk
>> _______________________________________________
>> Dros mailing list
>> Dros from net.bio.net
>> http://www.bio.net/biomail/listinfo/dros
>>
>
>

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