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Expression and Purification of P43 Toxoplasma gondii Surface Antigen

BACKGROUND: Toxoplasma gondii is an obligate intracellular protozoan parasite, capable of infecting all species of mammals including man. Congenital toxoplasmosis is more important during pregnancy for the first time. In this study we expressed and purified P43 Toxoplasma gondii tachyzoite and brady...

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Autores principales: Khanaliha, K, Motazedian, MH, Sarkari, B, Bandehpour, M, Sharifnia, Z, Kazemi, B
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Tehran University of Medical Sciences 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3469172/
https://www.ncbi.nlm.nih.gov/pubmed/23109962
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author Khanaliha, K
Motazedian, MH
Sarkari, B
Bandehpour, M
Sharifnia, Z
Kazemi, B
author_facet Khanaliha, K
Motazedian, MH
Sarkari, B
Bandehpour, M
Sharifnia, Z
Kazemi, B
author_sort Khanaliha, K
collection PubMed
description BACKGROUND: Toxoplasma gondii is an obligate intracellular protozoan parasite, capable of infecting all species of mammals including man. Congenital toxoplasmosis is more important during pregnancy for the first time. In this study we expressed and purified P43 Toxoplasma gondii tachyzoite and bradyzoite specific surface antigen. METHODS: The recombinant pGEMEX-1 contained Toxoplasma P43 coding sequence was transformed into E. coli and mass cultured in LB medium contained 100 μg/ml ampicillin at 37°C over night. The T7 promoter was induced by 1mM isopropyl-1-thio-ß-D-galactopyranoside (IPTG. Recombinant protein was purified by affinity chromatography and confirmed by gel diffusion dot blot and western blot,-using specific anti Toxoplasma antibodies. RESULTS: Recombinant plasmid was induced by IPTG and analyzed by SDS-PAGE. Recombinant protein was confirmed by Western-blot and dot blot using anti human Toxoplasma antibody. CONCLUSION: Recombinant Toxoplasma P43 was produced successfully.
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spelling pubmed-34691722012-10-29 Expression and Purification of P43 Toxoplasma gondii Surface Antigen Khanaliha, K Motazedian, MH Sarkari, B Bandehpour, M Sharifnia, Z Kazemi, B Iran J Parasitol Original Article BACKGROUND: Toxoplasma gondii is an obligate intracellular protozoan parasite, capable of infecting all species of mammals including man. Congenital toxoplasmosis is more important during pregnancy for the first time. In this study we expressed and purified P43 Toxoplasma gondii tachyzoite and bradyzoite specific surface antigen. METHODS: The recombinant pGEMEX-1 contained Toxoplasma P43 coding sequence was transformed into E. coli and mass cultured in LB medium contained 100 μg/ml ampicillin at 37°C over night. The T7 promoter was induced by 1mM isopropyl-1-thio-ß-D-galactopyranoside (IPTG. Recombinant protein was purified by affinity chromatography and confirmed by gel diffusion dot blot and western blot,-using specific anti Toxoplasma antibodies. RESULTS: Recombinant plasmid was induced by IPTG and analyzed by SDS-PAGE. Recombinant protein was confirmed by Western-blot and dot blot using anti human Toxoplasma antibody. CONCLUSION: Recombinant Toxoplasma P43 was produced successfully. Tehran University of Medical Sciences 2012 /pmc/articles/PMC3469172/ /pubmed/23109962 Text en © 2012 Iranian Society of Parasitology & Tehran University of Medical Sciences http://creativecommons.org/licenses/by/2.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Khanaliha, K
Motazedian, MH
Sarkari, B
Bandehpour, M
Sharifnia, Z
Kazemi, B
Expression and Purification of P43 Toxoplasma gondii Surface Antigen
title Expression and Purification of P43 Toxoplasma gondii Surface Antigen
title_full Expression and Purification of P43 Toxoplasma gondii Surface Antigen
title_fullStr Expression and Purification of P43 Toxoplasma gondii Surface Antigen
title_full_unstemmed Expression and Purification of P43 Toxoplasma gondii Surface Antigen
title_short Expression and Purification of P43 Toxoplasma gondii Surface Antigen
title_sort expression and purification of p43 toxoplasma gondii surface antigen
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3469172/
https://www.ncbi.nlm.nih.gov/pubmed/23109962
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