biostudies-literatureKiraly ONIDDK NIH HHS175-94https://www.ebi.ac.uk/biostudies/studies/S-EPMC3107599biostudies-literatureUnknown705Heterologously expressed proteins in Escherichia coli may undergo unwanted N-terminal processing by methionine and proline aminopeptidases. To overcome this problem, we present a system where the gene of interest is cloned as a fusion to a self-splicing mini-intein. This fusion construct is expressed in an engineered E. coli strain from which the pepP gene coding for aminopeptidase P has been deleted. We describe a protocol using human cationic trypsinogen as an example to demonstrate that recombinant proteins produced in this expression system contain homogeneous, unprocessed N-termini.Methods in molecular biology (Clifton, N.J.)Expression of recombinant proteins with uniform N-termini.PMC3107599R01 DK058088-06R01 DK058088-07R01 DK058088DK058088R01 DK058088-05A1Kiraly OGuan LSahin-Toth MfalseExpression of recombinant proteins with uniform N-termini.Heterologously expressed proteins in Escherichia coli may undergo unwanted N-terminal processing by methionine and proline aminopeptidases. To overcome this problem, we present a system where the gene of interest is cloned as a fusion to a self-splicing mini-intein. This fusion construct is expressed in an engineered E. coli strain from which the pepP gene coding for aminopeptidase P has been deleted. We describe a protocol using human cationic trypsinogen as an example to demonstrate that recombinant proteins produced in this expression system contain homogeneous, unprocessed N-termini.2011-01-01T00:00:00Z20112025-04-27T00:40:59.033Z2019-06-05T16:30:49ZS-EPMC31075992112538610.1007/978-1-61737-967-3_10