I am looking for a plasmid that can express my gene of interest without GFP fusion. But I want its selection marker like neomycin replaced by GFP or any other fluorescent protein so that I can select based on the fluorescence. Thank you in advance.
Hi:
you would find lots of ires-GFP plasmids could use. your GOI with CMV and the GFP with ires.
or you could fuse the GFP with your GOI by 2A peptide.
you also could cotransfection for example 1 ug of GOI and 10 ng of GFP, most GFP positive cells will have your GOI.
Junjie Shao. Not a fan of the IRES reporter system. It's great when it works and that takes some serious luck and planets to align. But I agree with you on T2A based constructs. They seem to be the best option yet.
pcDNA is a good plasmid. You can add the GFP sequence to it as a tag.
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