Author/Authors :
AGHDASI, Marjaneh Dept. of Parasitology - Faculty of Medical Sciences - University of Tarbiat Modarres, Tehran, Iran , GHAFFARIFAR, Fatemeh Dept. of Parasitology - Faculty of Medical Sciences - University of Tarbiat Modarres, Tehran, Iran , FOROOGHI, Fatemeh Dept. of Parasitology - Faculty of Medical Sciences - University of Tarbiat Modarres, Tehran, Iran , DALIMI ASL, Abdol Hossein Dept. of Parasitology - Faculty of Medical Sciences - University of Tarbiat Modarres, Tehran, Iran , SHARIFI, Zohre Research Center of Iranian Blood Transfusion Organization, Tehran, Iran , MASPI, Nahid Dept. of Parasitology - Faculty of Medicine - Ilam University of Medical Sciences, Ilam, Iran
Abstract :
Background: Toxoplasmosis is a common infection all around the world. During
pregnancy; it may lead to congenital disorders or abortion in human and animals.
Severe damage of toxoplasmosis indicates to require effective vaccine. One of
dense granules antigen is GRA4 that secrete from tachyzoite and bradyzoite. GRA4
genome is unique without intron and is one of the major immunogenic proteins
from Toxoplasma gondii.
Methods: We confirmed the cloning of GRA4 gene into pcDNA3 by restriction
enzyme and PCR of GRA4 gene with pcGRA4 plasmids as template. Then with
using calcium- phosphate method we transfected the pcGRA4 into CHO (Chinesehamster
ovary) cells. The yielded protein was separated by SDS-PAGE and
moved by electroblotting to nitrocellulose paper.
Results: Result of SDS-PAGE analysis showed the appearance of band approximately
42 kDa which was absent in the negative control, that was able to identify
toxoplasmosis antibody IgM+ serum in western blot analysis.
Conclusion: pcGRA4 plasmid is able to synthesis of antigenic protein in CHO
cells. The ability of pcGRA4 for induction of protective immune response against
toxoplasmosis will be evaluated in mouse model.
Keywords :
Toxoplasma gondii , GRA4 gene , Gene expression , CHO cell
