reporter gene fusion - (Jul/02/2007 )
Hey BioForum folks,
I'm trying to fuse a peptide to a reporter gene to use in mammalian cells( HEK293). I'm thinking about using β-galactosidase. So the lab has a β-galactosidase vector that can be used in Ecoli. I want to ask if this vector can be used in mammalian cells as well. My thinking is that i would not be able to use this vector because not only would the promoter not function in mammalian cells but the β-galactosidase sequence for mammalian cells is very different from the one in Ecoli (meaning I cant PCR out the β-galactosidase sequence and just insert after a mammalian promoter). Any ideas?
Also aside from β-galactosidase and GFP, what other reporter genes are good as fusion reporter genes in mammalian cells ?
So my lab has a luciferase reporter gene (PGL4.13 from promega). Do you know where I can get information and protocols to help me design a peptide-luciferase fusion protein? (things like what linker to use, ect).
We amplified the peptide and cloned it after the luciferase. We had 2 glycines after the luciferase without the stopcodon.
We used the pGL3-vector from Promega.