[Zbrafish] Re: CMV promoter coupled transgene expression

Tobias via zbrafish%40net.bio.net (by langenberg from gmail.com)
Wed Mar 10 03:12:15 EST 2010


Dear Santhosh,

while I personally never used CMV promoter for transgenesis, I believe
that it is by now well-known that it becomes silenced in successive
generations, maybe already in the founders. Which tissue do you want
your transgene to be expressed in? I would strongly recommend using a
zebrafish specific promoter with known expression profile and/or some
kind of inducible system like heatshock or Cre or Gal4/UAS. There is
no true ubiquitous promoter for zebrafish, to my knowledge. Happy
cloning!

Tobias

On Mar 9, 6:42 am, San <santhosh.kara... from gmail.com> wrote:
> We injected a CMV promoter coupled transgene using tol2 method. The
> founders died after first breeding. When we bred F1, the F2 are
> positive for the transgene by PCR. However we did not detected neither
> RNA nor hnRNA of the transgene using RT-PCR. Did anybody had this
> probelm before with CMV?
>
> Thanks.
> Santhosh.




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