Luciferase fusion protein? - (Aug/16/2013 )
I am planning to clone a firefly luciferase gene into an ORF of my system. I want to preserve the 5' RNA structure of the ORF. To do this I plan to have a fusion of the first 30 amino acids of the ORF and then the luciferase followed by a stop codon. My question is: is there precedence of luciferase functioning properly with ~30 amino acids fused to the N-terminus of the protein? Has anybody worked with luciferase fusion proteins? Certain proteins are functional with this many amino acids fused the N-terminus (e.g. Neomycin). I am just looking for feedback on whether or not luciferase can function as a fusion protein. Are C-terminal fusions OK?
OK, I asked around and found an answer.
A colleague has successfully fused 25 amino acids to the N-terminus of the firefly luciferase gene without affecting luciferase activity. I am also told there would not be any problems fusing the amino acids to the C-terminus as well.