I want to clone a protein from a gram-negative bacterium that has a natural periplasmic signal peptide. Is it ok to delete the signal sequence, put the N-terminal amino acid directly next to a methionine and clone it? Will this result in cytoplasmic expression? How might this affect the folding of the protein? Of these 3 ways to clone the protein, which is more likely to result in soluble expression -
1) deleting the signal sequence - cytoplasmic expression
2) cloning with the intact natural periplasmic signal peptide (will it be recognized by E.coli?)
3) deleting the signal sequence and placing the protein under pelB signal
I want to know if anyone has had experience of cloning such proteins. Is it possible to predict the outcome or does it depend on the protein?
Hi Avinash,
You can safey clone and express your protein without a signal sequence. This will indeed result in cytoplasmic expression without affecting correct folding. To summarise, I would suggest to go for the first option.
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