[Arabidopsis] Re: A question about studying plant gene subcellular localization byusing eYFP

Oliver Berkowitz via arab-gen%40net.bio.net (by oliver.berkowitz At anu.edu.au)
Mon Jan 29 18:01:55 EST 2007


Hi Kam,

had similar problems with my constructs (in pAVA vectors) changing to
another vector with a different linker between GFP and my cDNA worked. Also,
you might try transient expression by particle bombardment as you can
increase the amount of DNA you shoot and thus the expression of your
construct.

Cheers
Oliver

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Dr. Oliver Berkowitz
The Australian National University
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"Chan Kam Ho" <h0204121 At gmail.com> wrote in message
news:mailman.966.1170107703.19683.arab-gen At net.bio.net...
> Dear all,
>
> I have tried to study the subcellular localization of my gene by fusing it
> in frame with eYFP and also DsRed2. I have placed it under the control of
> CaMV and I have sequenced the constructs and I am sure that the construct
is
> correct and there is no misake in the construct. I tried to overexpress
the
> construct in transgenic Arabidopsis and since my construct also contains
> CaMV GUS I found that my transgenics are GUS positive but I can't see the
> fluorescence of eYFP/ DsRed2. Is it possible that the fluorescent proteins
> doesn't fold correctly when it is fused with my protein of interest and so
I
> can see the fluorescence? does anyone have similar experience to share?
What
> can I do now?
>
>
> Thanks in advance
> Kam Ho




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