title: Novel broad host range shuttle vectors for expression in Escherichia coli, Bacillus subtilis and Pseudomonas putida.
abstract: Novel shuttle vectors named pEBP were constructed to allow the gene expression in different bacterial hosts including Escherichia coli, Bacillus subtilis and Pseudomonas putida. These vectors share the inducible promoters P(T7) and P(Xyl) and a cos site to enable packaging of plasmid DNA into phage, and carry different multiple cloning sites and antibiotic resistance genes. Vector pEBP41 generally replicates episomally while pEBP18 replicates episomally in Gram-negative bacteria only, but integrates into the chromosome of B. subtilis. Plasmid copy numbers determined for E. coli and P. putida were in the range of 5-50 per cell. The functionality of pEBP18 and pEBP41 was confirmed by expression of two lipolytic enzymes, namely lipase A from B. subtilis and cutinase from the eukaryotic fungus Fusarium solani pisi in three different host strains. Additionally, we report here the construction of a T7 RNA polymerase-based expression strain of P. putida.
authors.affiliation: Institute of Molecular Enzyme Technology, Heinrich-Heine-University Duesseldorf, Research Center Juelich, D-52426 Juelich, Germany.
authors.forename: Sonja Christina
authors.initials: SC
authors.lastname: Troeschel
authors.forename: Stephan
authors.initials: S
authors.lastname: Thies
authors.forename: Olga
authors.initials: O
authors.lastname: Link
authors.forename: Catherine Isabell
authors.initials: CI
authors.lastname: Real
authors.forename: Katja
authors.initials: K
authors.lastname: Knops
authors.forename: Susanne
authors.initials: S
authors.lastname: Wilhelm
authors.forename: Frank
authors.initials: F
authors.lastname: Rosenau
authors.forename: Karl-Erich
authors.initials: KE
authors.lastname: Jaeger
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D005670:Fusarium / Q000201:enzymology / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C019952:cutinase; D008049:Lipase