Construction of pOGOduet - An inducible, bicistronic vector for synthesis of recombinant proteins in Corynebacterium...

The Gram-positive Corynebacterium glutamicum is widely known for its application in the industrial production of amino acids and as a non-pathogenic model organism for cell wall biosynthesis in the group of CMN bacteria. For biotechnological and physiological studies often co-expression of recombinant genes is required, however for C. glutamicum no vector for the independent co-expression of two genes was described. We here created the novel expression vector pOGOduet for C. glutamicum, which carries the ColE1 replicon of E. coli and the pBL1 replicon of C. glutamicum and two independently inducible promoters Ptac and Ptet each followed by unique multiple cloning sites. Functionality of pOGOduet is tested by coexpression of genes for the fluorescent proteins eCFP and mVenus; fluorescence of the reporters varies in dependence of the inducer concentrations present in the culture broth. These experiments demonstrate that the vector pOGOduet fulfills the task for individually inducible expression of two genes of interest in C. glutamicum.