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:: Volume 15, Issue 3 (Quarterly 2011) ::
Feyz 2011, 15(3): 194-199 Back to browse issues page
Determination and cloning of the gene encoding EG95 protein in Iranian isolate of Echinococcus granulosus
Shahabedin Sarvi, Abdolhossein Dalimi-Asl *, Fatemeh Ghaffarifar, Zohreh Sharifi
Tarbiat Modares University , dalimi_a@modares.ac.ir
Abstract:   (8376 Views)

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.

Keywords: Echinococcus granulosus, EG95 gene, Protoscoleces, pTZ57R/T plasmid
Full-Text [PDF 383 kb]   (2267 Downloads)    
Type of Study: Research | Subject: General
Received: 2011/09/10 | Published: 2011/09/15
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Sarvi S, Dalimi-Asl A, Ghaffarifar F, Sharifi Z. Determination and cloning of the gene encoding EG95 protein in Iranian isolate of Echinococcus granulosus. Feyz. 2011; 15 (3) :194-199
URL: http://feyz.kaums.ac.ir/article-1-1238-en.html

Volume 15, Issue 3 (Quarterly 2011) Back to browse issues page
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