A method to export gene products to the growth medium of Gram negative bacteria.

摘要

A process for expressing proteins in Gram(-) bacteria and providing for extracellular secretion thereof, comprising the steps:
<UnorderedLists id="ula01" listStyle="none"><ListItem>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;</ListItem><ListItem>b) cultivating the bacterium under conditions resulting in filamentous growth; and</ListItem><ListItem>c) 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; and
proteins obtained by such process.</ListItem></UnorderedLists>