General expression vectors for Staphylococcus carnosus enabled efficient production of the outer membrane protein A of Klebsiella pneumoniae.
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.
Hansson
Marianne
M
Department of Biotechnology, SCFAB, Royal Institute of Technology (KTH), SE-10691, Stockholm, Sweden.
Samuelson
Patrik
P
Nguyen
Thien Ngoc
TN
Ståhl
Stefan
S
D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D015151:Immunoblotting; D007711:Klebsiella pneumoniae / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013210:Staphylococcus / Q000235:genetics*
D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins; C078167:OMPA outer membrane proteins