So, I would like to create a fusion protein where my GFP is C-terminally tagged to my proteins. However, the restriction enzyme site on this vector has HindIII site on the 5' end where I want to put my gene in and there are several HindiIII sites in my genes. There is limited enzyme sites in that location so one must use HindIII. If one used another enzyme, which is 20 or so bp downstream, I would have the additional 20 bps (and the corresponding aa) in my protein and I don't want that.
I looked up to see if HindIII shares compatible ends with other enzymes but it doesn't. I have thought about using homologous recombination (Clonetech Infusion) but they have had success with a limited number of vectors and I don't know if mine would work.
What other ways can I insert my gene into that particular region? HELP!
Submit your paper to J Biol Methods today!
Restriction Enzyme sites in my genes
1 reply to this topic