Rapid orientated cloning in a shuttle vector allowing modulated gene expression in Bacillus subtilis

An expression vector for systematic protein overproduction in Bacillus subt ilis has been constructed. It derives from pDG148 and combines the main pro perty or this vector, i.e. conditional expression of the gene in response t o isopropyl beta -D-thiogalactopyranoside, with (i) rapid orientated clonin g by a ligation independent procedure and (ii) a ribosome binding site of h igh translational efficiency. When used for overproduction of several prote ins in B. subtilis, this vector gave good levels of protein synthesis. (C) 2001 Federation of European Microbiological Societies. Published by Elsevie r Science B.V. All rights reserved.