A library of Treponema pallidum genomic DNA fragments produced by partial Sau3A digestion was established in Escherichia coli K12 using the plasmid vector pAT153. The library was screened using immune syphilitic rabbit serum and six recombinant phenotypes expressing eight treponemal polypeptides were detected. With two exceptions, all the recombinant gene products were the same size as polypeptides detected on Western immunoblots of T. pallidum. The genes encoding three novel gene products, with molecular masses in SDS-PAGE of 42, 17 and 15·5 kDa, which had not been cloned previously from T. pallidum were also identified. Monoclonal antibodies which reacted with four of the eight recombinant polypeptides were generated.
BaileyM.J.,
CockayneA.,
PennC.W.1987; Monoclonal antibodies directed against surface- associated polypeptides of Treponemapallidum define a biologically active antigen.. Journal of General Microbiology 133:1793–1803
BeallJ.A.,
MitchellG.F.1986; Identification of a particular antigen from a parasite cDNA library using antibodies affinity purified from selected portions of Western blots.. Journal of Immunological Methods 86:217–233
BrahamshaB.,
GreenbergE.P.1988; Immuno- electron microscope analysis of Spirochaetaaurantiaperiplasmic flagella and molecular cloning of a major axial filament antigen.. Abstracts of the Annual Meeting of the American Society for Microbiology p. 193:
BroomeW.S.,
GilbertW.1978; Immunological screening method to detect specific translation products.. Proceedings of the National Academy of Sciences of the United States of America 75:2746–2749
CockayneA.,
BaileyM.J.,
PennC.W.1987; Analysis of sheath and core structures of the axial filament of Treponema pallidum.. Journal of General Microbiology 133:1397–1407
Van EmbdenJ.D.,
Van Der DonkH.J.,
Van EijkR.V.,
Van Der HeideH.G.,
De JongJ.A.,
Van OlderenM.F.,
OsterhausA.D.,
SchoulsL.M.1983; Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12.. Infection and Immunity 42:187–196
FehnigerT.E.,
WalfieldA.M.,
CunninghamT.M.,
RadolfJ.D.,
MillerJ.N.,
LovettM.A.1984; Purification and characterization of a cloned protease-resistant Treponema pallidumspecific antigen.. Infection and Immunity 46:598–607
HanffP.A.,
FehnigerT.E.,
MillerJ.N.,
LovettM.A.1982; Humoral immune response in human syphilis to polypeptides of Treponema pallidum.. Journal of Immunology 129:1287–1291
HanffP.A.,
BishopN.H.,
MillerJ.N.,
LovettM.A.1983b; Humoral immune response in experimental syphilis to polypeptides of Treponema pallidum.. Journal of Immunology 131:1973–1977
HansenE.B.,
PedersenP.E.,
SchoulsL.M.,
SeverinE.,
Van EmbdenJ.D.A.1985; Genetic characterization and partial sequence determination of a Treponema pallidum operon expressing two immunogenic membrane proteins in Escherichia coli.. Journal of Bacteriology 162:1227–1237
HenselU.,
Jobst-WellensiekH.,
BhakdiS.1985; Sodium dodecyl sulphate-polyacrylamide gel electrophoresis immunoblotting as a serological tool in the diagnosis of syphilitic infections.. Journal of Clinical Microbiology 21:82–87
HinderssonP.,
BangsdorfJ.1987; Cloning and expression of 4 Treponema pallidum antigens identified by monoclonal antibodies.. Abstracts of the Annual Meeting of the American Society for Microbiology D47 p. 79:
HinderssonP.,
KnudsenJ.D.,
AxelsenN.H.1987; Cloning and expression of Treponema pallidum common antigen (Tp-4) in Escherichia coli K12.. Journal of General Microbiology 133:587–596
HsuP.-L.,
QinM.,
NorrisS.J.,
SellS.1988; Isolation and characterization of recombinant Escherichia coli clones secreting a 24-kilodalton antigen of Treponema pallidum.. Infection and Immunity 56:1135–1143
IjsselmuidenO.E.,
SchoulsL.M.,
StolzE.,
AelbersG.N.M.,
AgterbergC.M.,
Van EmbdenJ.D.A.1989; Sensitivity and specificity of an enzyme linked immunosorbent assay using the recombinant DNA-derived Treponema pallidum protein TmpA for serodiagnosis of syphilis and the potential use of TmpA for assessing the effect of antibiotic therapy.. Journal of Clinical Microbiology 27:152–157
JonesS.A.,
MarchittoK.S.,
MillerJ.N.,
NorgardM.A.1984; Monoclonal antibody with haemagglutination, immobilization, and neutralization activities defines an immunodominant, 47,000 mol. wt., surface-exposed immunogen of Treponema pallidum (Nichols).. Journal of Experimental Medicine 160:1404–1420
LukehartS.A.,
Baker-ZanderS.A.,
GubishE.R.1982; Identification of Treponema pallidumantigens: comparison with a non-pathogenic tre- poneme.. Journal of Immunology 129:833–838
NorgardM.V.,
ChamberlainN.R.,
SwancuttM.A.,
GoldbergM.S.1986; Cloning and expression of the major 47-kilodalton surface immunogen of Treponema pallidum in Escherichia coli.. Infection and Immunity 54:500–506
NorgardM.V.,
MillerJ.N.1983; Cloning and expression of Treponema pallidum (Nichols) antigen genes in Escherichia coli.. Infection and Immunity 42:435–445
O’ConnorC.G.,
AshmanL.K.1982; Application of nitrocellulose transfer techniques and alkaline phosphatase-conjugated anti-immunoglobulin for determination of the specificity of monoclonal antibodies to protein mixtures.. Journal of Immuno-logical Methods 54:267–271
PedersenN.S.,
AxelsenN.H.,
JorgensenB.B.,
PetersenC.S.1980; Antibodies in secondary syphilis against five of forty Reiter treponeme antigens.. Scandinavian Journal of Immunology 11:629–633
PennC.W.1983; The use of the rabbit as a model for syphilis infection.. In Experimental Bacterial and Parasitic Infections pp. 145–151KeuschG.,
WadstromT.
Edited by Amsterdam: Elsevier Biomedical;
PennC.W.,
CockayneA.,
BaileyM.J.1985; The outer membrane of Treponema pallidum: biological significance and biochemical properties.. Journal of General Microbiology 131:2349–2357
PetersonK.M.,
BasemanJ.B.,
AldereteJ.F.1986; Isolation of a Treponema pallidum gene encoding immunodominant outer envelope protein P6, which reacts with sera from patients at different stages of syphilis.. Journal of Experimental Medicine 164:1160–1170
RadolfJ.D.,
NorgardM.V.1988; Pathogen specificity of T. pallidum subsp. pallidum membrane proteins identified by phase partitioning with Triton X-114.. Infection and Immunity 56:1825–1828
SanchezP.J.,
MccrackenG.H.JrWendelG.D.,
OlsenK.,
ThrelkeldN.,
NorgardM.V.1989; Molecular analysis of the fetal IgM response to Treponema pallidum antigens: implications for improved serodiagnosis of congenital syphilis.. Journal of Infectious Diseases 159:508–517
StrugnellR.A.,
WilliamsW.F.,
DrummondL.P.,
PedersenJ.S.,
TohB.H.,
FaineS.1986; Development of increased seroblot reactivity against a 45,000 dalton polypeptide of Treponema pallidum(Nichols) correlates with the establishment of chancre immunity in syphilitic rabbits.. Infection and Immunity 51:957–960
SwancuttM.A.,
TwehousD.A.,
NorgardM.V.1986; Monoclonal antibody selection and analysis of a recombinant DNA-derived surface immunogen of Treponema pallidum expressed in Escherichia coli.. Infection and Immunity 52:110–119