Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus
Background: Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection that occurs by the larval stages of taeniid cestodes of the genus Echinococcus. Iran is known as endemic region for this infection in the world. Vaccination has been considered as a good prevention method for this d...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2012-06-01
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| Series: | Iranian Journal of Parasitology |
| Subjects: | |
| Online Access: | http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/20951.pdf&manuscript_id=20951 |
| _version_ | 1818350805026078720 |
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| author | F Ghafarifar A Dalimi S Sarvi |
| author_facet | F Ghafarifar A Dalimi S Sarvi |
| author_sort | F Ghafarifar |
| collection | DOAJ |
| description | Background: Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection that occurs by the larval stages of taeniid cestodes of the genus Echinococcus. Iran is known as endemic region for this infection in the world. Vaccination has been considered as a good prevention method for this disease. Recombinant vaccines containing EG95 protein, against E. granulosus, has shown a high degree of protection against E. granulosus infection. In this study EG95 gene was extracted from Iranian isolates of E. granulosus and then cloned and expressed in expression vector.Methods: Protoscoleces were collected from sheep hydatid cysts. Then DNA and RNA were extracted from protoscoleces, and amplified by PCR and RT-PCR with specific primer. Afterward the purified RT-PCR products were successfully ligated into pTZ57R/T plasmid vector. The pcDNA3 plasmid was used as expression vector and Eg95 fragment sub cloned into this plasmid. The pcEG95 plasmid was digested by restriction enzymes to confirm cloning of this gene in pcDNA3 plasmid. In last step, the subcloned gene was expressed in CHO as eukaryotic cell.Results: EG95 fragment successfully was subcloned in pcDNA3 and EG95 protein was expressed by eukaryotic cell. The recombinant EG95 protein was confirmed by SDS-PAGE and Western blot.Conclusion: Recombinant plasmid of pcEG95 was constructed successfully and express of recombinant EG95 protein was confirmed. |
| first_indexed | 2024-12-13T18:27:40Z |
| format | Article |
| id | doaj.art-b68ed7b0bd3343359cf5bdb70520a991 |
| institution | Directory Open Access Journal |
| issn | 1735-7020 2008-238X |
| language | English |
| last_indexed | 2024-12-13T18:27:40Z |
| publishDate | 2012-06-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Parasitology |
| spelling | doaj.art-b68ed7b0bd3343359cf5bdb70520a9912022-12-21T23:35:33ZengTehran University of Medical SciencesIranian Journal of Parasitology1735-70202008-238X2012-06-017217Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus GranulosusF GhafarifarA DalimiS SarviBackground: Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection that occurs by the larval stages of taeniid cestodes of the genus Echinococcus. Iran is known as endemic region for this infection in the world. Vaccination has been considered as a good prevention method for this disease. Recombinant vaccines containing EG95 protein, against E. granulosus, has shown a high degree of protection against E. granulosus infection. In this study EG95 gene was extracted from Iranian isolates of E. granulosus and then cloned and expressed in expression vector.Methods: Protoscoleces were collected from sheep hydatid cysts. Then DNA and RNA were extracted from protoscoleces, and amplified by PCR and RT-PCR with specific primer. Afterward the purified RT-PCR products were successfully ligated into pTZ57R/T plasmid vector. The pcDNA3 plasmid was used as expression vector and Eg95 fragment sub cloned into this plasmid. The pcEG95 plasmid was digested by restriction enzymes to confirm cloning of this gene in pcDNA3 plasmid. In last step, the subcloned gene was expressed in CHO as eukaryotic cell.Results: EG95 fragment successfully was subcloned in pcDNA3 and EG95 protein was expressed by eukaryotic cell. The recombinant EG95 protein was confirmed by SDS-PAGE and Western blot.Conclusion: Recombinant plasmid of pcEG95 was constructed successfully and express of recombinant EG95 protein was confirmed.http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/20951.pdf&manuscript_id=20951Echinococcus GranulosusEG95PcDNA3 Plasmid |
| spellingShingle | F Ghafarifar A Dalimi S Sarvi Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus Iranian Journal of Parasitology Echinococcus Granulosus EG95 PcDNA3 Plasmid |
| title | Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus |
| title_full | Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus |
| title_fullStr | Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus |
| title_full_unstemmed | Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus |
| title_short | Molecular Cloning and Expression of EG95 Gene of Iranian Isolates of Echinococcus Granulosus |
| title_sort | molecular cloning and expression of eg95 gene of iranian isolates of echinococcus granulosus |
| topic | Echinococcus Granulosus EG95 PcDNA3 Plasmid |
| url | http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/20951.pdf&manuscript_id=20951 |
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