Ghaffarifar, Fatemeh, Nordin, Rahme, Sharifi, Zohreh, Dalimi, Abdolhossein, Roudbar Mohammadi, Shahla, Ghasemi Nikoo, Sakineh (2008). Cloning and sequencing of granular antigen7 (GRA7) in TOPO vector and transformation in TOP10. , 11(0), 73-80.
Fatemeh Ghaffarifar; Rahme Nordin; Zohreh Sharifi; Abdolhossein Dalimi; Shahla Roudbar Mohammadi; Sakineh Ghasemi Nikoo. "Cloning and sequencing of granular antigen7 (GRA7) in TOPO vector and transformation in TOP10". , 11, 0, 2008, 73-80.
Ghaffarifar, Fatemeh, Nordin, Rahme, Sharifi, Zohreh, Dalimi, Abdolhossein, Roudbar Mohammadi, Shahla, Ghasemi Nikoo, Sakineh (2008). 'Cloning and sequencing of granular antigen7 (GRA7) in TOPO vector and transformation in TOP10', , 11(0), pp. 73-80.
Ghaffarifar, Fatemeh, Nordin, Rahme, Sharifi, Zohreh, Dalimi, Abdolhossein, Roudbar Mohammadi, Shahla, Ghasemi Nikoo, Sakineh Cloning and sequencing of granular antigen7 (GRA7) in TOPO vector and transformation in TOP10. , 2008; 11(0): 73-80.

Cloning and sequencing of granular antigen7 (GRA7) in TOPO vector and transformation in TOP10

Pathobiology Reserach
Article 10, Volume 11, Issue 0, 2008, Pages 73-80    PDF (2.12 M)
Authors
Fatemeh Ghaffarifar* 1; Rahme Nordin2; Zohreh Sharifi3; Abdolhossein Dalimi4; Shahla Roudbar Mohammadi5; Sakineh Ghasemi Nikoo6
1Associated Professor, Department of Parasitology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
2Professor, Research Center of Molecular Medicine, USM University, Penang, Malaysia
3Assistant Professor, Research Center of Virology, Iranian Blood Transfusion Organization, Tehran, Iran
4Professor, Department of Parasitology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
5Assistant Professor, Department of Mycology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
6M.Sc., Department of Parasitology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objective: Toxoplasmosis, caused by an intracellular protozoan parasite, and the Toxoplasma gondii, is widespread throughout the world. In recent years, significant progress has been made in the identification of vaccine candidates which could induce a protective response.
GRA7, an excretory 29 kDa Toxoplasma gondii a dense granular antigen released by infected host cells. In tachyzoite-infected cells, p29 accumulates within the parasitophorous vacuole and co-localizes with its delimiting membrane.
Materials and Methods: In the present work, first genomic DNA of Toxoplasma gondii was extracted and used for amplifying of GRA7 gene as a template. Then PCR product was extracted from agarose gel and cloned into TOPO vector. The plasmid containing GRA7 gene was extracted from the transformed bacteria (TOP10 strain) and sequenced.
Results: Sequence analysis of GRA7 gene cloned into TOPO vector showed only one base difference when composed with the gene bank sequence for RH strain was only one base.
Conclusion: The results indicated that this clone is suitable for subcloning in Prokaryotic and Eukaryotic plasmid.
Keywords
Toxoplasma gondii; Cloning; Sequencing; GRA7 gene
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