Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7
Abstract
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:
-
- Bellport, NY
- San Antonio, TX
- Issue Date:
- Research Org.:
- Associated Universities, Inc., Upton, NY (United States)
- OSTI Identifier:
- 870674
- Patent Number(s):
- 5571718
- Application Number:
- 07/941,523
- Assignee:
- Associated Universities, Inc. (Washington, DC)
- Patent Classifications (CPCs):
-
A - HUMAN NECESSITIES A61 - MEDICAL OR VETERINARY SCIENCE A61K - PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES
- DOE Contract Number:
- AC02-76CH00016
- Resource Type:
- Patent
- 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/
Citation Formats
Dunn, John J, and Barbour, Alan G. Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7. United States: N. p., 1996.
Web.
Dunn, John J, & Barbour, Alan G. Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7. United States.
Dunn, John J, and Barbour, Alan G. Tue .
"Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7". United States. https://www.osti.gov/servlets/purl/870674.
@article{osti_870674,
title = {Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7},
author = {Dunn, John J and Barbour, Alan G},
abstractNote = {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.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {1996},
month = {11}
}
Works referenced in this record:
Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins
journal, April 1990
- Brandt, M. E.; Riley, B. S.; Radolf, J. D.
- Infection and Immunity, Vol. 58, Issue 4
Protection of mice against the Lyme disease agent by immunizing with recombinant OspA
journal, October 1990
- Fikrig, E.; Barthold, S.; Kantor, F.
- Science, Vol. 250, Issue 4980
Purification and characterization of the outer membrane lipoprotein from an Escherichia coli mutant altered in the signal sequence of prolipoprotein.
journal, February 1980
- Lin, J. J.; Kanazawa, H.; Wu, H. C.
- Journal of Biological Chemistry, Vol. 255, Issue 3
A single recombinant plasmid expressing two major outer surface proteins of the Lyme disease spirochete
journal, February 1985
- Howe, T.; Mayer, L.; Barbour, A.
- Science, Vol. 227, Issue 4687
Juxtaposition of expressed variable antigen genes with a conserved telomere in the bacterium Borrelia hermsii.
journal, August 1990
- Kitten, T.; Barbour, A. G.
- Proceedings of the National Academy of Sciences, Vol. 87, Issue 16