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

Patent ·
OSTI ID:870674
 [1];  [2]
  1. Bellport, NY
  2. San Antonio, TX
+ Show Author Affiliations

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.

Research Organization:
Associated Universities, Inc., Upton, NY (United States)
DOE Contract Number:
AC02-76CH00016
Assignee:
Associated Universities, Inc. (Washington, DC)
Patent Number(s):
US 5571718
Application Number:
07/941,523
OSTI ID:
870674
Country of Publication:
United States
Language:
English

References (5)

Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins journal April 1990
Protection of mice against the Lyme disease agent by immunizing with recombinant OspA journal October 1990
Purification and characterization of the outer membrane lipoprotein from an Escherichia coli mutant altered in the signal sequence of prolipoprotein. journal February 1980
A single recombinant plasmid expressing two major outer surface proteins of the Lyme disease spirochete journal February 1985
Juxtaposition of expressed variable antigen genes with a conserved telomere in the bacterium Borrelia hermsii. journal August 1990

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cloning
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preparing
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oligonucleotide
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/435/530/999/