pGC310

Model Organism: Worms Methods & Reagents
1

Hi,
I am planning to amplify m-cherry from pGC310 plasmid to fuse it to my gene of interest by PCR sowing. I was wondering if this would give me optimum reporter expression since I do not plan to include any sequence upstream of the m-cherry coding sequence since I do not want NLS sequence which is present upstream of m-cherry sequence?I just want to make sure that I do not get rid of any additional sequence that might be required for proper expression of the transgene.
Thanks for help

2

I think that should be fine. I’m not aware of any sequence upstream of mCherry required for expression. I suppose it depends on what you’re trying to do. Is this a transcriptional reporter? Translational fusion? If so, N- or C-terminal? For translational fusions, it could always be good to leave a linker of flexible amino acids (serines and glycines) between the tag and the protein to reduce chances of disrupting your protein of interest.

3

Thanks for your response. I am trying to make a C-terminal translational fusion