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:: Volume 17, Issue 1 (Bimonthly 2013) ::
Feyz 2013, 17(1): 8-13 Back to browse issues page
Characterization and expression of GRA7 gene of Toxoplasma gondii RH strain in eukaryotic pcDNA3 plasmid
Hosein Vazini, Fatemeh Ghaffarifar *, Zohreh Sharifi, Abdolhosein Dalimi-Asl
Tarbiat Modares University , ghafarif@modares.ac.ir
Abstract:   (5282 Views)

Background: Toxoplasmosis, as a widespread and important disease, can cause severe complications in the congenital and HIV cases. Vaccination is one of the preventive approaches in humans and domestic animals. The GRA7, an excretory 29 kDa Toxoplasma gondii dense granule antigen released by the infected host cells , has been considered as a candidate to produce a vaccine .

Materials and Methods: In this experimental study, the inserted plasmid containing GRA7 gene was extracted from TOP10 bacteria and digested with the BamH1 and EcoR1 enzymes. The isolated gene was inserted into the pcDNA3 plasmid . The cloning was confirmed by the PCR and sequencing. The protein expression was confirmed by the SDS-PAGE and Western blotting.

Results: The results showed that the GRA7 gene was cloned into the pcDNA3 plasmid. The isolated gene cloned in pcDNA3 was confirmed by PCR and showed the 733bp band. The pcGRA7 plasmid expressed in the CHO cells showed the 29 kDa band using the SDS-PAGE and Western blotting.

Conclusion: Considering that cloning of GRA7 gene has been done in the pcDNA3 plasmid and transformed in the eukaryotic cells, it can be used for the DNA vaccine researches.

 

Keywords: Toxoplasma gondii RH strain, Cloning, pc-GRA7, Eukaryotic pcDNA3 plasmid
Full-Text [PDF 696 kb]   (1209 Downloads)    
Type of Study: Research | Subject: medicine, paraclinic
Received: 2013/03/4 | Accepted: 2013/03/16 | Published: 2013/03/16
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Vazini H, Ghaffarifar F, Sharifi Z, Dalimi-Asl A. Characterization and expression of GRA7 gene of Toxoplasma gondii RH strain in eukaryotic pcDNA3 plasmid. Feyz. 2013; 17 (1) :8-13
URL: http://feyz.kaums.ac.ir/article-1-1839-en.html


Volume 17, Issue 1 (Bimonthly 2013) Back to browse issues page
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