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

A method is provided 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. 38 figs.
Inventors:
;
Issue Date:
OSTI Identifier:
392642
Assignee:
Associated Universities, Inc., Washington, DC (United States) PTO; SCA: 550200; PA: EDB-96:167742; SN: 96001682407
Patent Number(s):
US 5,571,718/A/
Application Number:
PAN: 7-941,523
Contract Number:
AC02-76CH00016
Resource Relation:
Other Information: PBD: 5 Nov 1996
Research Org:
Associated Universities Inc
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; LIPOPROTEINS; PROTEIN ENGINEERING; BACTERIA; GENETIC ENGINEERING; BIOTECHNOLOGY; DNA-CLONING; GENE AMPLIFICATION; RECOMBINANT DNA; ESCHERICHIA COLI