| abstract |
The present invention refers to an expression vector (A) for enhanced expression of a protein of interest encoding a fusion protein comprising at least a secretory signal sequence (A1); a solubilizing agent (A2); a proteolytic cleavage site (A3); and a protein of interest (A4), optionally a tag for purification of the protein of interest (A5) and optionally a heterologously expressed protease (A6), capable of cleaving the proteolytic cleavage site (A3) subsequent to or during secretion of the fusion protein. Alternatively, the protease (A6) may be endogenously expressed in the host cell, may be provided to the host cell using a second expression vector (B) and/or may be added extracellularly. The present invention furthermore provides methods for using the inventive expression vectors (A) and, optionally, ( B ), in vitro and in vivo, and kits, comprising these expression vectors and/or cells comprising same. |