Hello all,

I am struggling to expressed a codon optimised paramecium fused proteins in HEK293 cell line. The two proteins are naturally nuclei proteins, however, one of the fused protein shuttle between nuclear and cytoplasm. I added 1 NLS at the C-terminal of the fused protein, but the protein was unable to be translocated to the nucleus.

How can I increase the nuclear translocation of the protein? What could be the reason for the protein not being able to enter the nucleus? Size? nuclear export sequences? How can I find them and may be mutate them?

Best regards

Elvis