help finding eukaryote expression vector - (Aug/08/2007 )
I am looking for a vector which i can use to express a mouse protein in eukaryote (mamalian) cells.
to complicate matters I want to add a his tag for purification, which i want to remove after purification.
so the His tag has t be N terminal (the protein also has an n-terminal signal peptide)
I was thinking of CMV HIS(removable)/gene
Can anybody tell me what would be a suitable vector (and perhaps donate me a tube?)
you may try the pIRESneo vector. CMV promoter, an ires and selection marker. It's up to you to clone appropriate removable Tag maintaining a MCS. It's feasible easily with 2 annealed oligos, and may serve for future genes.
Otherwise, i know that the [beta]globin intron may be inserted for enhancing the maturation of the transgene mRNA.
Check pcDNA4 His (invitrogen). It has HIS in N terminus. We use pcDNA3.1+ , but the HIS is in the C terminus.