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Enhancer GFP construct: what about the "frame"?


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#1 tobikenobi

tobikenobi

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Posted 12 January 2011 - 12:44 AM

Hello everybody,

I have a question that bugs me, however, it might be a really stupid one:

I am planning to generate a transgenic mouse line that expresses GFP under control of a tissue specific enhancer sequence.
So, I made me some primers flanking the enhancer sequence with RE sites, amplified, and exchanged the promoter from my GFP vector with the enhancer sequence.
My question now is: Does the enhancer sequence need to be in a certain spacing regarding the start of my GFP? Is there a frame that cannot be compromised?

The ability of this sequence to direct GFP expression to specific parts of the neural system has been described (PMID 20171205), however, in the methods part, the only info is that they cloned the enhancer sequence into a gfp expression cassette. How does such a cassette typically look like in regard to the inserted sequence and the start of the GFP?

Thank you so much for your help!

Best regards,

Tobi




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