I am trying to find a promoter, to make reporter line for few genes but the genes are not well studied, so no information available. Is there any resources to find promoters for zebrafish gene?
Hi Palsamy,
the way people actually do is, if somebody has already done it, you ask for the p5E plasmid for the tol2 system.
If it hasn't been done yet, then you should get a BAC that contains your gene (you can find these in ZFIN.ORG), tag your gene of interest (or replace it with GFP), add the tol2 sites and then use a transposase to insert it in the genome. After that, you would screen for the ones that give you GFP signal.
Hope this helps! Good luck,
J.
Hi Joaquin,Thank you
Thank you.
I am taking 4 kb upstream seq and fusing to GFP. Hopefully that works.
People have used in mice, so i am using those length and features.
Thanks.
yes usually a 3kb upstream region would suffice. Just seuence to check for the presence of all the usual promoter elements.
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