Determination and cloning of the gene encoding EG95 protein in Iranian isolate of Echinococcus granulosus

Sarvi, Sh. and Dalimi-Asl, A. and Ghaffarifar, F. and Sharifi, Z. (2011) Determination and cloning of the gene encoding EG95 protein in Iranian isolate of Echinococcus granulosus. Feyz Journal of Kashan University of Medical Sciences, 15.

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Official URL: http://feyz.kaums.ac.ir/article-1-1238-en.html
DOI: UNSPECIFIED

Abstract

Background: Echinococcus granulosus is a cestode parasite that causes cystic hydatid disease in humans worldwide. The gene encoding EG95 protein may be a good candidate to design a DNA vaccine to prevent the disease. Considering the importance of EG95 gene and the scarceness of research on it in Iran, this study was carried out to determine and clone the gene encoding EG95 from Iranian isolate of E. granulosus. Materials and Methods: At the first stage, protoscoleces was isolated from hydatid cyst fluid and then RNA was extracted from protoscoleces and after performing RT-PCR, the amplified cDNA samples were detected by gel electrophoresis. In next stage, the obtained gene was cloned in pTZ57R/T vector. Two methods were used for conformation of cloning: colony PCR amplification and digestion with the EcoRI and XhoI restriction enzymes. Finally, the cloned EG95 gene in pTZ57R/T vector was sequenced.Results: Homological comparison of sequences showed that cDNA of EG95 in Iranian isolate of E. granulosus had 492 bp and was different from the standard strain of EG95 reported from New Zealand and Australia (X90928.1). Moreover, cloning of EG95 gene in pTZ57R/T plasmid was confirmed by digestion of this plasmid with the restriction enzymes. Conclusion: The EG95 gene was cloned in pTZ57R/T plasmid successfully and this plasmid can be used to design a DNA vaccine in further studies.

Item Type: Article
Subjects: Immunology and Microbiology
Divisions: Feyz journal
Depositing User: ART . editor
Date Deposited: 10 May 2017 12:05
Last Modified: 25 May 2017 13:26
URI: http://eprints.kaums.ac.ir/id/eprint/1912

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