KLONIERUNG UND EXPRESSION VON BORRELIA LIPOPROTEINEN

• Main Authors: BARBOUR, ALAN G, DUNN, JOHN J
• Format: Patent
• Language: German
• Published: 15.08.2001
• Edition: 7
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; HUMAN NECESSITIES; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

A method is provided herein for preparing highly expressed and soluble recombinant variations of Borrelia lipoproteins such as Borrelia burgdorferi outer surface protein A (OspA). The method includes synthesizing a set of oligonucleotides for a portion of the OspA gene excluding a region coding for a recognition site for lipoprotein signal peptidase II, amplifying the template DNA utilizing the polymerase chain reaction, purifying the amplification products, cloning the amplification products into a suitable expression vector, transforming a suitable host utilizing the cloned expression vector, cultivating the transformed host for protein production and subsequently isolating and purifying the resulting protein. Also provided are soluble, recombinant variations of Borrelia burgdorferi outer surface protein A (OspA) as well as the cloned pET9 plasmid, pET9-OspA. Transformed E. coli host BL21(DE3)/pLysS, pET9-OspA, is also disclosed.