Molecular characterization and expression of p23 (OspC) from a North American strain of Borrelia burgdorferi.
We have found that sera from patients with early stages of Lyme disease contain predominant immunoglobulin M reactivity to a major 23-kDa protein (p23) from Borrelia burgdorferi 2591 isolated in Connecticut. To characterize this immunodominant antigen, we cloned and sequenced p23 and found it to be 83% identical by nucleotide sequence and 75% identical by amino acid sequenced to pC (recently renamed OspC), an abundantly expressed protein on the outer surface of PKo, a European strain of B. burgdorferi (B. Wilske, V. Preac-Mursic, S. Jauris, A. Hofmann, I. Pradel, E. Soutschek, E.Schwab, G. Will, and G. Wanner, Infect. Immun. 61:2182-2191, 1993). In addition, immunoelectron microscopy localized p23 to the outer membrane, confirming that p23 is the strain 2591 homolog of OspC. The North American strain B31, commonly used in serologic assays for Lyme disease, does not express OspC. Northern (RNA) blot analysis detected low levels of ospC mRNA in B31, and DNA sequencing of the ospC gene from B31 revealed a 54-bp deletion in the upstream regulatory region, possibly accounting for the low transcriptional activity of ospC. The ospC coding region from B31 was cloned and antibody-reactive OspC was expressed in Escherichia coli. An immunoglobulin M enzyme-linked immunosorbent assay using recombinant OspC as the target antigen shows promise for the serodiagnosis of early stages of Lyme disease.
Padula
S J
SJ
Department of Medicine, University of Connecticut Health Center, Farmington 06030-1310.
Sampieri
A
A
Dias
F
F
Szczepanski
A
A
Ryan
R W
RW
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015748:Borrelia burgdorferi Group / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015870:Gene Expression; D005798:Genes, Bacterial; D006801:Humans; D007075:Immunoglobulin M / Q000096:biosynthesis; D008193:Lyme Disease / Q000175:diagnosis / Q000276:immunology / Q000382:microbiology; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009656:North America; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D007075:Immunoglobulin M; C079616:OspC protein; D011993:Recombinant Fusion Proteins