I am expressing my protein in a baculovirus vector. After expression I find most of my protein in the cell and some of it is secreted into the medium.
SOSUI predicted that my protein has a signal peptide. I was interested to add a signal peptide so that most of the protein is secreted.
Questions:
1. Do I need to cleave the native signal peptide before adding an insect specific signal peptide?
2. Which signal peptide would be the best
a)Honey bee melettin b) GP67 signal peptidec)SP1
Sona. it depends on where you want your protein to end up. if you are happy for it to remain in the cell and purify it from there then you should take your protein signal peptide off and ensure you provide a start codon for your protein. this way your protein should accumulate intracellularly.
If however you wish your protein to be secreted then you should replace your signal peptide sequence (I am assuming you are working with a non-insect protein) with the honey bee mellitin sequence
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