TY - JOUR T1 - Characterization and expression of GRA7 gene of Toxoplasma gondii RH strain in eukaryotic pcDNA3 plasmid TT - شناسایی و بیان ژنGRA7 توکسوپلاسما گونده‌ای سویه RH در پلاسمید یوکاریوت بیانی pcDNA3 JF - KAUMS JO - KAUMS VL - 17 IS - 1 UR - http://feyz.kaums.ac.ir/article-1-1839-en.html Y1 - 2013 SP - 8 EP - 13 KW - Toxoplasma gondii RH strain KW - Cloning KW - pc-GRA7 KW - Eukaryotic pcDNA3 plasmid N2 - 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.   M3 ER -