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Title: Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7

A method is provided herein for preparing soluble recombinant variations of Borrelia lipoproteins such as Borrelia burgdorferi outer surface protein A (OspA) and outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The method includes synthesizing a set of oligonucleotide primers, amplifying the template DNA utilizing the PCR, 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), outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The expression vectors harboring DNA encoding the recombinant variations, pET9-OspA, pET9-OspB and pET9-Vmp7, as well as the E. coli host BL21(DE3)/pLysS transformed with each of these vectors, are also disclosed.
Inventors:
 [1];  [2]
  1. (Bellport, NY)
  2. (San Antonio, TX)
Issue Date:
OSTI Identifier:
870674
Assignee:
Associated Universities, Inc. (Washington, DC) BNL
Patent Number(s):
US 5571718
Application Number:
07/941,523
Contract Number:
AC02-76CH00016
Research Org:
ASSOC UNIVERSITIES INC
Country of Publication:
United States
Language:
English
Subject:
cloning; expression; soluble; truncated; variants; borrelia; ospa; ospb; vmp7; method; provided; preparing; recombinant; variations; lipoproteins; burgdorferi; outer; surface; protein; hermsii; variable; major; synthesizing; set; oligonucleotide; primers; amplifying; template; dna; utilizing; pcr; purifying; amplification; products; suitable; vector; transforming; host; cloned; cultivating; transformed; production; subsequently; isolating; resulting; vectors; harboring; encoding; pet9-ospa; pet9-ospb; pet9-vmp7; coli; bl21; de3; plyss; disclosed; suitable host; outer surface; dna encoding; transformed host; expression vectors; oligonucleotide primers; expression vector; oligonucleotide primer; dna utilizing; /435/530/999/