Cloning and expression of borrelia lipoproteins

Details Cloning and expression of borrelia lipoproteins Cloning and expression of borrelia lipoproteins

C - Chemistry, Metallurgy

12

N

C12N 15/31 (2006.01) C07H 21/04 (2006.01) C07K 14/20 (2006.01) C12N 1/21 (2006.01) C12N 15/67 (2006.01) C12N 15/70 (2006.01) A61K 38/00 (2006.01) A61K 39/00 (2006.01)

Patent

CA 2057536

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 oligonucleotide primers, 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.

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