Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue

Darrin R. Akins, Stephen F. Porcella, Taissia G. Popova, Dmitriy Shevchenko, Scott I. Baker, Minyue Li, Michael V. Norgard, Justin D. Radolf

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Abstract

Protein export signals from the low-passage 297 strain of Borrelia burgdorferi were cloned as fusions with an Escherichia coli alkaline phosphatase (PhoA) reporter lacking a signal sequence. One PhoA+ clone (BbK2.10-PhoA) was derived from a borrelial lipoprotein. Although the polypeptide encoded by the full-length bbk2.10 gene had 76% similarity and 56% identity to outer surface protein F (OspF) from B. burgdorferi strain N40, antibodies directed against recombinant forms of the two proteins revealed that they were not cross-reactive. The nucleotide sequences of bbk2.10 and ospF from the N40 and 297 strains, respectively, were determined to confirm that the N40 and 297 strains each contained both genes. Southern blot analysis revealed that bbk2.10 is a single-copy gene and that the B. burgdorferi strain 297 and N40 genomes appeared to contain one other gene more closely related to ospF than bbk2.10. It was particularly noteworthy that ospF, but not bbk2.10, was expressed in vitro while B. burgdorferi-infected mice generated antibodies reactive with both lipoproteins. To help confirm that the BbK2.10-reactive antibodies produced by the B. burgdorferi-infected mice were specific for that protein, a second gene, bbk2.11, which hybridized with the ospF probe was cloned; the corresponding polypeptide reacted strongly with OspF antisera but failed to react with BbK2.10-specific antisera. Taken together, these data demonstrate that BbK2.10, BbK2.11, and OspF comprise a B. burgdorferi lipoprotein family and that at least one member (BbK2.10) appears to be expressed only during infection.

Original languageEnglish (US)
Pages (from-to)507-520
Number of pages14
JournalMolecular Microbiology
Volume18
Issue number3
StatePublished - Nov 1995

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Borrelia burgdorferi
Membrane Proteins
Lipoproteins
Genes
Phosphoric Monoester Hydrolases
Antibodies
Immune Sera
Peptides
Protein Sorting Signals
Southern Blotting
Alkaline Phosphatase
Proteins
Clone Cells
In Vitro Techniques
Genome
Escherichia coli
Infection

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Akins, D. R., Porcella, S. F., Popova, T. G., Shevchenko, D., Baker, S. I., Li, M., ... Radolf, J. D. (1995). Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue. Molecular Microbiology, 18(3), 507-520.

Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue. / Akins, Darrin R.; Porcella, Stephen F.; Popova, Taissia G.; Shevchenko, Dmitriy; Baker, Scott I.; Li, Minyue; Norgard, Michael V.; Radolf, Justin D.

In: Molecular Microbiology, Vol. 18, No. 3, 11.1995, p. 507-520.

Research output: Contribution to journalArticle

Akins, DR, Porcella, SF, Popova, TG, Shevchenko, D, Baker, SI, Li, M, Norgard, MV & Radolf, JD 1995, 'Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue', Molecular Microbiology, vol. 18, no. 3, pp. 507-520.
Akins DR, Porcella SF, Popova TG, Shevchenko D, Baker SI, Li M et al. Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue. Molecular Microbiology. 1995 Nov;18(3):507-520.
Akins, Darrin R. ; Porcella, Stephen F. ; Popova, Taissia G. ; Shevchenko, Dmitriy ; Baker, Scott I. ; Li, Minyue ; Norgard, Michael V. ; Radolf, Justin D. / Evidence for in vivo but not in vitro expression of a Borrelia burgdorferi outer surface protein F (OspF) homologue. In: Molecular Microbiology. 1995 ; Vol. 18, No. 3. pp. 507-520.
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abstract = "Protein export signals from the low-passage 297 strain of Borrelia burgdorferi were cloned as fusions with an Escherichia coli alkaline phosphatase (PhoA) reporter lacking a signal sequence. One PhoA+ clone (BbK2.10-PhoA) was derived from a borrelial lipoprotein. Although the polypeptide encoded by the full-length bbk2.10 gene had 76{\%} similarity and 56{\%} identity to outer surface protein F (OspF) from B. burgdorferi strain N40, antibodies directed against recombinant forms of the two proteins revealed that they were not cross-reactive. The nucleotide sequences of bbk2.10 and ospF from the N40 and 297 strains, respectively, were determined to confirm that the N40 and 297 strains each contained both genes. Southern blot analysis revealed that bbk2.10 is a single-copy gene and that the B. burgdorferi strain 297 and N40 genomes appeared to contain one other gene more closely related to ospF than bbk2.10. It was particularly noteworthy that ospF, but not bbk2.10, was expressed in vitro while B. burgdorferi-infected mice generated antibodies reactive with both lipoproteins. To help confirm that the BbK2.10-reactive antibodies produced by the B. burgdorferi-infected mice were specific for that protein, a second gene, bbk2.11, which hybridized with the ospF probe was cloned; the corresponding polypeptide reacted strongly with OspF antisera but failed to react with BbK2.10-specific antisera. Taken together, these data demonstrate that BbK2.10, BbK2.11, and OspF comprise a B. burgdorferi lipoprotein family and that at least one member (BbK2.10) appears to be expressed only during infection.",
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AU - Akins, Darrin R.

AU - Porcella, Stephen F.

AU - Popova, Taissia G.

AU - Shevchenko, Dmitriy

AU - Baker, Scott I.

AU - Li, Minyue

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AU - Radolf, Justin D.

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AB - Protein export signals from the low-passage 297 strain of Borrelia burgdorferi were cloned as fusions with an Escherichia coli alkaline phosphatase (PhoA) reporter lacking a signal sequence. One PhoA+ clone (BbK2.10-PhoA) was derived from a borrelial lipoprotein. Although the polypeptide encoded by the full-length bbk2.10 gene had 76% similarity and 56% identity to outer surface protein F (OspF) from B. burgdorferi strain N40, antibodies directed against recombinant forms of the two proteins revealed that they were not cross-reactive. The nucleotide sequences of bbk2.10 and ospF from the N40 and 297 strains, respectively, were determined to confirm that the N40 and 297 strains each contained both genes. Southern blot analysis revealed that bbk2.10 is a single-copy gene and that the B. burgdorferi strain 297 and N40 genomes appeared to contain one other gene more closely related to ospF than bbk2.10. It was particularly noteworthy that ospF, but not bbk2.10, was expressed in vitro while B. burgdorferi-infected mice generated antibodies reactive with both lipoproteins. To help confirm that the BbK2.10-reactive antibodies produced by the B. burgdorferi-infected mice were specific for that protein, a second gene, bbk2.11, which hybridized with the ospF probe was cloned; the corresponding polypeptide reacted strongly with OspF antisera but failed to react with BbK2.10-specific antisera. Taken together, these data demonstrate that BbK2.10, BbK2.11, and OspF comprise a B. burgdorferi lipoprotein family and that at least one member (BbK2.10) appears to be expressed only during infection.

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