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AssigneeNascaCell Technologies AG
InventorsHarald Kolmar, Andreas Christmann, Alexander Wentzel
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The inventive method allows peptides or polypeptides to be exposed on the surface of gram-negative host bacteria using specific intimin-based anchor modules. Intimins with shortened carboxy terminals have been found to be particularly suitable anchor modules for passenger domains in the exterior E.coli cell membrane. According to said method, host bacteria are transformed using vectors, on which are located a fused nucleic acid sequence consisting of a sequence segment which codes for an intimin with a shortened carboxy terminal and a nucleic acid sequence segment which codes for the passenger peptide that is to be exposed. The invention permits a particularly large number of passenger domains to be exposed on the cell surface of the bacteria, without adversely affecting the viability of the bacteria.
Process for exposing peptides and proteins on the surface of host bacteria, in which one
Process for producing a variant population of surface-exposed peptides/polypeptides/proteins and for identifying bacteria, which carry the peptides/polypeptides/proteins with a desired property, in which the process has the following steps:
Expression vector for the expression of a fusion gene under the control of an exogeneously inducible promoter in which there is, in operative linkage with the promoter, a coding sequence for a desired passenger peptide in the continuous reading frame downstream to a coding sequence for an Intimin shortened by at least the D3 domain in the carboxy-terminal region of 280 amino acids s the anchoring domain for the passenger peptide
Dependencies are inferred from the claim text.