Direkt zum Inhalt
Merck
  • Toxoplasma gondii: cloning, expression and immunoreactivity of recombinant ROP5 and ROP18 antigens.

Toxoplasma gondii: cloning, expression and immunoreactivity of recombinant ROP5 and ROP18 antigens.

Experimental parasitology (2015-01-17)
Marcin M Grzybowski, Bożena Dziadek, Jarosław Dziadek, Justyna Gatkowska, Katarzyna Dzitko, Henryka Długońska
ZUSAMMENFASSUNG

Early diagnosis and determining the infective stage are critical for effective therapy of toxoplasmosis. Owing to the progress in biotechnology, commonly used native, non-standardized diagnostic antigens should be replaced by genetically engineered antigens. The recombinant proteins are also promising components of subunit vaccines against Toxoplasma gondii infections. A strategic biological role of rhoptry proteins (ROP) in parasitophorous vacuole biogenesis and virulence of the parasite creates a necessity for an intensive study on the serological activity and immunogenicity of newly developed recombinant ROP antigens. Our findings indicate that all generated preparations of recombinant ROP5 and ROP18 antigens, expressed in Escherichia coli bacteria, are recognized by specific antibodies produced during acute and chronic infections in inbred laboratory mice. We noticed, for the first time, that ROP5 IgM antibodies are an early and sensitive marker of T. gondii infection. The proven immunoreactivity of the obtained preparations has become a premise for a further study on their utility in routine diagnosis of human and animal toxoplasmosis as well as in the immunoprevention of T. gondii infection (as the main or supplementary component of the vaccine).

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Wasserstoffperoxid -Lösung, contains inhibitor, 30 wt. % in H2O, ACS reagent
Sigma-Aldrich
Wasserstoffperoxid -Lösung, 30 % (w/w) in H2O, contains stabilizer
Sigma-Aldrich
2-Mercaptoethanol, Molecular Biology, suitable for electrophoresis, suitable for cell culture, BioReagent, 99% (GC/titration)
Sigma-Aldrich
2-Mercaptoethanol, ≥99.0%
Sigma-Aldrich
Phenylmethansulfonylfluorid, ≥98.5% (GC)
Sigma-Aldrich
Wasserstoffperoxid -Lösung, 50 wt. % in H2O, stabilized
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
2-Mercaptoethanol, BioUltra, Molecular Biology, ≥99.0% (GC)
Sigma-Aldrich
Ampicillin, anhydrous, 96.0-102.0% (anhydrous basis)
Millipore
Wasserstoffperoxid -Lösung, 3%, suitable for microbiology
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, ≥99%
Supelco
Wasserstoffperoxid -Lösung, ≥30%, for trace analysis
Sigma-Aldrich
Phenylmethansulfonylfluorid, ≥99.0% (T)
Sigma-Aldrich
Wasserstoffperoxid -Lösung, contains inhibitor, 30 wt. % in H2O, meets USP testing specifications
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, ≥98% (TLC)
Sigma-Aldrich
Isopropyl β-D-1-Thiogalactopyranosid, ≥99% (TLC)
Sigma-Aldrich
Wasserstoffperoxid -Lösung, 34.5-36.5%
Sigma-Aldrich
Isopropyl-β-D-thiogalactopyranosid -Lösung, ReadyMade IPTG solution for Blue-white screening
Supelco
Wasserstoffperoxid -Lösung, 30 % (w/w), for ultratrace analysis
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, ≥98.0% (NT)
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidin, tablet, 1 mg substrate per tablet
Supelco
Ampicillin
SAFC
Isopropyl β-D-1-Thiogalactopyranosid
Sigma-Aldrich
Ampicillin, meets USP testing specifications
Supelco
2-Mercaptoethanol, derivatization grade (HPLC), LiChropur, ≥99.0% (GC)
USP
Ampicillin, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Wasserstoffperoxid -Lösung, tested according to Ph. Eur.
Ampicillin, anhydrous, European Pharmacopoeia (EP) Reference Standard
Supelco
3,3′,5,5′-Tetramethylbenzidin, standard for GC