Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein

A Hemphill, R Felleisen, Bernadette Connolly, B Gottstein, B Hentrich, N Müller

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

Neospora caninum is an apicomplexan parasite of veterinary importance which invades many different cell types and tissues. N. caninum tachyzoites proliferate intracellularly by endodyogeny. Eventually the massive proliferation of tachyzoites leads to host cell lysis and the newly formed parasites are released and invade neighbouring cells. Tachyzoite cell surface molecules could serve as ligands, mediating host cell adhesion and invasion. Nc-p43 is a recently identified N. caninum tachyzoite surface protein which is functionally involved in the processes leading to host cell invasion in vitro. Affinity-purified antibodies directed against Nc-p43 were used to screen a lambda gt22A-cDNA expression library constructed from N. caninum tachyzoites. The cDNA insert of one immunoreactive clone was subcloned and expressed in E. coli as a poly-histidine fusion protein. The identity of the resulting recombinant antigen termed recNc-p43 was confirmed by immunoblotting, immunofluorescence and electron microscopy using affinity-purified antibodies. The sequence of the cDNA insert encoding recNc-p43 was determined. Analysis of the deduced amino acid sequence revealed that Nc-p43 exhibited similarity to SAG1 (p30) and SAG3 (p43), 2 major surface antigens of Toxoplasma gondii tachyzoites. These similarities were not reflected on the immunochemical level, since no cross-antigenicity between SAG1, SAG3 and Nc-p43 was observed.
Original languageEnglish
Pages (from-to)581-90
Number of pages10
JournalParasitology
Volume115 ( Pt 6)
Publication statusPublished - 1 Dec 1997

Fingerprint

Neospora
tachyzoites
Neospora caninum
surface proteins
Membrane Proteins
Complementary DNA
Clone Cells
clones
Antibody Affinity
cell invasion
Parasites
cells
Protein Sequence Analysis
Toxoplasma
recombinant antigens
parasites
Surface Antigens
Gene Library
Fluorescence Microscopy
Immunoblotting

Keywords

  • Amino Acid Sequence
  • Animals
  • Antigens, Protozoan
  • Antigens, Surface
  • Cercopithecus aethiops
  • DNA Transposable Elements
  • DNA, Complementary
  • DNA, Protozoan
  • Fluorescent Antibody Technique
  • Gene Library
  • Host-Parasite Interactions
  • Immunoblotting
  • Immunohistochemistry
  • Membrane Glycoproteins
  • Membrane Proteins
  • Molecular Sequence Data
  • Neospora
  • Protozoan Proteins
  • Toxoplasma
  • Vero Cells

Cite this

Hemphill, A., Felleisen, R., Connolly, B., Gottstein, B., Hentrich, B., & Müller, N. (1997). Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein. Parasitology, 115 ( Pt 6), 581-90.

Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein. / Hemphill, A; Felleisen, R; Connolly, Bernadette; Gottstein, B; Hentrich, B; Müller, N.

In: Parasitology, Vol. 115 ( Pt 6), 01.12.1997, p. 581-90.

Research output: Contribution to journalArticle

Hemphill, A, Felleisen, R, Connolly, B, Gottstein, B, Hentrich, B & Müller, N 1997, 'Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein', Parasitology, vol. 115 ( Pt 6), pp. 581-90.
Hemphill A, Felleisen R, Connolly B, Gottstein B, Hentrich B, Müller N. Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein. Parasitology. 1997 Dec 1;115 ( Pt 6):581-90.
Hemphill, A ; Felleisen, R ; Connolly, Bernadette ; Gottstein, B ; Hentrich, B ; Müller, N. / Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein. In: Parasitology. 1997 ; Vol. 115 ( Pt 6). pp. 581-90.
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abstract = "Neospora caninum is an apicomplexan parasite of veterinary importance which invades many different cell types and tissues. N. caninum tachyzoites proliferate intracellularly by endodyogeny. Eventually the massive proliferation of tachyzoites leads to host cell lysis and the newly formed parasites are released and invade neighbouring cells. Tachyzoite cell surface molecules could serve as ligands, mediating host cell adhesion and invasion. Nc-p43 is a recently identified N. caninum tachyzoite surface protein which is functionally involved in the processes leading to host cell invasion in vitro. Affinity-purified antibodies directed against Nc-p43 were used to screen a lambda gt22A-cDNA expression library constructed from N. caninum tachyzoites. The cDNA insert of one immunoreactive clone was subcloned and expressed in E. coli as a poly-histidine fusion protein. The identity of the resulting recombinant antigen termed recNc-p43 was confirmed by immunoblotting, immunofluorescence and electron microscopy using affinity-purified antibodies. The sequence of the cDNA insert encoding recNc-p43 was determined. Analysis of the deduced amino acid sequence revealed that Nc-p43 exhibited similarity to SAG1 (p30) and SAG3 (p43), 2 major surface antigens of Toxoplasma gondii tachyzoites. These similarities were not reflected on the immunochemical level, since no cross-antigenicity between SAG1, SAG3 and Nc-p43 was observed.",
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T1 - Characterization of a cDNA-clone encoding Nc-p43, a major Neospora caninum tachyzoite surface protein

AU - Hemphill, A

AU - Felleisen, R

AU - Connolly, Bernadette

AU - Gottstein, B

AU - Hentrich, B

AU - Müller, N

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N2 - Neospora caninum is an apicomplexan parasite of veterinary importance which invades many different cell types and tissues. N. caninum tachyzoites proliferate intracellularly by endodyogeny. Eventually the massive proliferation of tachyzoites leads to host cell lysis and the newly formed parasites are released and invade neighbouring cells. Tachyzoite cell surface molecules could serve as ligands, mediating host cell adhesion and invasion. Nc-p43 is a recently identified N. caninum tachyzoite surface protein which is functionally involved in the processes leading to host cell invasion in vitro. Affinity-purified antibodies directed against Nc-p43 were used to screen a lambda gt22A-cDNA expression library constructed from N. caninum tachyzoites. The cDNA insert of one immunoreactive clone was subcloned and expressed in E. coli as a poly-histidine fusion protein. The identity of the resulting recombinant antigen termed recNc-p43 was confirmed by immunoblotting, immunofluorescence and electron microscopy using affinity-purified antibodies. The sequence of the cDNA insert encoding recNc-p43 was determined. Analysis of the deduced amino acid sequence revealed that Nc-p43 exhibited similarity to SAG1 (p30) and SAG3 (p43), 2 major surface antigens of Toxoplasma gondii tachyzoites. These similarities were not reflected on the immunochemical level, since no cross-antigenicity between SAG1, SAG3 and Nc-p43 was observed.

AB - Neospora caninum is an apicomplexan parasite of veterinary importance which invades many different cell types and tissues. N. caninum tachyzoites proliferate intracellularly by endodyogeny. Eventually the massive proliferation of tachyzoites leads to host cell lysis and the newly formed parasites are released and invade neighbouring cells. Tachyzoite cell surface molecules could serve as ligands, mediating host cell adhesion and invasion. Nc-p43 is a recently identified N. caninum tachyzoite surface protein which is functionally involved in the processes leading to host cell invasion in vitro. Affinity-purified antibodies directed against Nc-p43 were used to screen a lambda gt22A-cDNA expression library constructed from N. caninum tachyzoites. The cDNA insert of one immunoreactive clone was subcloned and expressed in E. coli as a poly-histidine fusion protein. The identity of the resulting recombinant antigen termed recNc-p43 was confirmed by immunoblotting, immunofluorescence and electron microscopy using affinity-purified antibodies. The sequence of the cDNA insert encoding recNc-p43 was determined. Analysis of the deduced amino acid sequence revealed that Nc-p43 exhibited similarity to SAG1 (p30) and SAG3 (p43), 2 major surface antigens of Toxoplasma gondii tachyzoites. These similarities were not reflected on the immunochemical level, since no cross-antigenicity between SAG1, SAG3 and Nc-p43 was observed.

KW - Amino Acid Sequence

KW - Animals

KW - Antigens, Protozoan

KW - Antigens, Surface

KW - Cercopithecus aethiops

KW - DNA Transposable Elements

KW - DNA, Complementary

KW - DNA, Protozoan

KW - Fluorescent Antibody Technique

KW - Gene Library

KW - Host-Parasite Interactions

KW - Immunoblotting

KW - Immunohistochemistry

KW - Membrane Glycoproteins

KW - Membrane Proteins

KW - Molecular Sequence Data

KW - Neospora

KW - Protozoan Proteins

KW - Toxoplasma

KW - Vero Cells

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VL - 115 ( Pt 6)

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EP - 590

JO - Parasitology

JF - Parasitology

SN - 0031-1820

ER -