i have a protein that is not naturally secreted out of the cell..so i decided to add a secretory signal (eg. IL-2 signal sequence) at 5' end of the gene...
kozak-secretory signal-my gene of interest...
my question is --- whether secretory signal should be in continuation with my gene or there can be any amino acid in between....[(GCCACC(kozak)IL-2signal sequenceATGCAA(my gene)]...
There might be additional sequence between your protein of interest and signal sequence.
Ideally to avoid influence of secondary structure whenever adding a foreign sequence to a new protein, I add a "linker sequence"of amino acids that has a proline glycine motif that would break the bend or structure and not influence the folding of the native part of the protein, thereby allowing full activity. So yes, i would recommend you add a linker.
Olga and subhojit,
please see the below link...i am planning similarly to my protein
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC27539
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