abstract |
A process for expressing proteins in Gram(-) bacteria and providing for extracellular secretion thereof, comprising the steps:a) introducing into a Gram(-) bacterium a recombinant DNA construction comprising a promoter, a signal sequence enabling translocation and processing, and a structural gene encoding the desired protein to be expressed;b) cultivating the bacterium under conditions resulting in filamentous growth; andc) recovering the extracellularly secreted protein;a recombinant DNA construction comprising:a promoter, a signal sequence and a structural gene including a cleavage region,wherein the structural gene is of the formula:(E)<sub>n</sub>(B)<sub>m</sub> - Y,where n is an integer ≧ 1, m is an integer, Y is a DNA sequence encoding a desired protein and E and B are the genes corresponding to the protein A regions E and B, with the proviso that the structural gene is different from that encoding natural protein A;a plasmid vector comprising such recombinant DNA construction;a Gram(-) bacterium harbouring said recombinant DNA construction; andproteins obtained by such process. |