Novel broad host range shuttle vectors for expression in Escherichia coli, Bacillus subtilis and Pseudomonas putida

作者：Troeschel Sonja Christina; Thies Stephan; Link Olga; Real Catherine Isabell; Knops Katja; Wilhelm Susanne; Rosenau Frank; Jaeger Karl Erich^*

来源：Journal of Biotechnology, 2012, 161(2): 71-79.

DOI：10.1016/j.jbiotec.2012.02.020

• Broad host range shuttle vector
• Enzyme expression
• Lipase

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.

• 出版日期2012-10-15