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Keywords:

  • pBBR1 origin;
  • PBAD promoter;
  • plasmids;
  • mobilizable vector

Abstract

Expression of recombinant proteins across a range of different host organisms is often necessary in metabolic engineering applications. Doing so can be facilitated by the use of vectors having origins of replication with a broad-host-range, the option for antibiotic resistance cassettes that are compatible with a particular host, in addition to sequences allowing for the effective transcription and translation of target proteins. We have created a modular set of broad-host-range expression vectors for protein expression in Gram negative bacteria. These vectors use the broad-host-range pBBR1 replicon as well as the arabinose-inducible PBAD promoter and are available with six different antibiotic resistance cassettes. We have demonstrated the use of these vectors in Escherichia coli, Pseudomonas putida, and Burkholderia cepacia. Biotechnol. Bioeng. 2010;106: 326–332. © 2010 Wiley Periodicals, Inc.