2 replies to this topic

[Abrahahm Hu]

Hi all
I am constructing a vector---Promoter::GFP-protein
The protein will be fuctioned as a marker gene together with GFP.
My question is whether it is necessary to add the ATG after the GFP coding sequence and 3'UTR of the protein to the end of the protein coning sequence.
Thanks very much!

[bob1]

Abrahahm Hu, on Jul 11 2010, 10:42 PM, said:

My question is whether it is necessary to add the ATG after the GFP coding sequence and 3'UTR of the protein to the end of the protein coning sequence.

It may or may not be important to have the starting Met of the protein - it may affect folding in some fashion if it is not there. However, having a whacking great tag like GFP in front of the protein will probably affect it in some fashion anyway.

You don't need the UTR as far as I know.

[Rsm]

Yes, better get rid of the UTR asap.
Well, it depends if you want to see degradation of your mRNA, or translational inhibition... but I guess you don't.

Cheers,

Minna