Volume 24, Issue 112 (5-2014)
J Mazandaran Univ Med Sci 2014, 24(112): 42-49 |
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Ahmadpour E, Sarvi S, Daryani A, Sharif M, Hashemi Souteh M, Mizani A et al . Cloning and sequencing the plasmid encoding dense granule antigen 14 (GRA14) of Toxoplasma gondii RH strain. J Mazandaran Univ Med Sci 2014; 24 (112) :42-49
URL: http://jmums.mazums.ac.ir/article-1-3038-en.html
URL: http://jmums.mazums.ac.ir/article-1-3038-en.html
Ehsan Ahmadpour 
, Shahabeddin Sarvi , Ahmad Daryani , Mehdi Sharif , Mohammad-Bagher Hashemi Souteh , Azadeh Mizani , Kian Rezaee
, Shahabeddin Sarvi , Ahmad Daryani , Mehdi Sharif , Mohammad-Bagher Hashemi Souteh , Azadeh Mizani , Kian Rezaee
Abstract: (10901 Views)
Background and purpose: Toxoplasmosis is a common parasitic disease throughout the world and one-third of the population has antibodies to Toxoplasma gondii. This disease causes serious medical problems in fetuses and immunocompromised individuals. As gene encoding protein GRA14 can be considered as a suitable target for DNA vaccine and designing diagnostic kits the aim of this study was to do cloning and sequencing the gene encoding GRA14 protein of Toxoplasma gondii RH strain.
Materials and methods: DNA extraction was performed on harvested tachyzoites from mouse peritoneal fluid, then PCR carried out and amplification products were analyzed by gel electrophoresis. GRA14 gene was cloned in pTG19-T as a cloning vector, then recombinant plasmid confirmed by the colony-PCR and restriction enzyme digestion using HindIII and EcoRI, followed by sequencing.
Results: Evaluation of PCR products by agarose gel electrophoresis and analysis of nucleotide sequencing of 1227 bp gene encoding the protein GRA14, revealed the complete homology with the recorded sequences in the gene bank. Furthermore, cloning of GRA14 gene in pTG19-T vector was confirmed with colony PCR and restriction enzyme digestion.
Conclusion: The results showed that the GRA14 gene was successfully cloned into the pTG19-T vector and this plasmid can be used to design DNA vaccines and diagnostic kits in further studies.
Type of Study: Research(Original) |
Subject:
parasitology
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