Rapid orientated cloning in a shuttle vector allowing modulated gene expression in Bacillus subtilis.
An expression vector for systematic protein overproduction in Bacillus subtilis has been constructed. It derives from pDG148 and combines the main property of this vector, i.e. conditional expression of the gene in response to isopropylbeta-D-thiogalactopyranoside, with (i) rapid orientated cloning by a ligation independent procedure and (ii) a ribosome binding site of high translational efficiency. When used for overproduction of several proteins in B. subtilis, this vector gave good levels of protein synthesis.
Joseph
P
P
Laboratoire de Chimie Bactérienne, Institut de Biologie Structurale et Microbiologie, CNRS 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 02, France.
Fantino
J R
JR
Herbaud
M L
ML
Denizot
F
F
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008969:Molecular Sequence Data; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D007544:Isopropyl Thiogalactoside