Method to export gene products to the growth medium of gram negative bacteria

US5156959A · kind A · utility

Assignee

• KabiGen AB · SE

Inventors

• Lars Abrahmsen · Bromma, SE
• Tomas Moks · Täby, SE
• Bjorn Nilsson · Foster City, US
• Mathias Uhlen · Uppsala, SE

Key dates

• Filing date: Oct 24, 1990
• Grant date: Oct 20, 1992
• Expiry date: Oct 24, 2010

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2319/705
• WIPO field: Biotechnology
• WIPO sector: Chemistry

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; and c) recovering the extracellularly secreted protein; a recombinant DNA construction comprising: PA1 a promoter, a signal sequence and a structural gene including a cleavage region, PA0 wherein the structural gene is of the formula: EQU (E).sub.n (B).sub.m -Y, PA0 where n is an integer .gtoreq.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; PA1 a plasmid vector comprising such recombinant DNA construction; PA1 a Gram(-) bacterium harbouring said recombinant DNA construction; and PA1 proteins obtained by such process.