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General expression vectors for Staphylococcus carnosus enabled efficient production of the outer membrane protein A of Klebsiella pneumoniae
Authors:Hansson Marianne  Samuelson Patrik  Nguyen Thien Ngoc  Ståhl Stefan
Institution:Department of Biotechnology, SCFAB, Royal Institute of Technology (KTH), SE-10691, Stockholm, Sweden.
Abstract:General expression vectors, designed for intracellular expression or secretion of recombinant proteins in the non-pathogenic Staphylococcus carnosus, were constructed. Both vector systems encode two different affinity tags, an upstream albumin binding protein and a downstream hexahistidyl peptide, and are furnished with cleavage sites for two site-specific proteases for optional affinity tag removal. To evaluate the novel vectors, the gene encoding the outer membrane protein A (OmpA) of Klebsiella pneumoniae was introduced into the vectors. Efficient production was demonstrated in both systems, although, as expected for OmpA fusions, somewhat better intracellularly, and the fusion proteins could be recovered as full-length products by affinity chromatography.
Keywords:Albumin binding protein  Intracellular expression  Outer membrane protein A  Secretion              Klebsiella pneumoniae                        Staphylococcus carnosus
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