fusion protein construction - (Oct/03/2009 )
I am going to construct a gusion protein between an enzyme (alkaline phosphatase) and a neomycine resitance gene for expression in mammalian cells, I study in some articles that in some cases a linker sequence is added between two proteins, please let me know if:
1. when create a fusion protein what is the risk that individual proteins loss their activity
2. is adding a linker necessary and what kind of linker works best
I was doing similar kind of work as you, and my constructs involved GFP + a protein of interest at the C terminus of the EGFP construct. I had to include a linker, 15 amino acid, rich in glycine and serine and that did the job, although we did observe a reduction in the brighnest of EGFP-fusion construct vs EGFP by itself. Hope this information helps.