Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

Expression of Plasmid Encoded GRA4 Gene of Toxoplasma gondii RH Strain in CHO Eukaryotic Cells

Sat May 25 21:43:08 2024

(2018) Expression of Plasmid Encoded GRA4 Gene of Toxoplasma gondii RH Strain in CHO Eukaryotic Cells. Iranian Journal of Parasitology. pp. 392-398. ISSN 1735-7020

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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.

Item Type: Article
Creators:
CreatorsEmail
Aghdasi, M.UNSPECIFIED
Ghaffarifar, F.UNSPECIFIED
Forooghi, F.UNSPECIFIED
Dalimi Asl, A. H.UNSPECIFIED
Sharifi, Z.UNSPECIFIED
Maspi, N.UNSPECIFIED
Keywords: Toxoplasma gondii GRA4 gene Gene expression CHO cell orally infected mice parasitophorous vacuole antigens DNA apicomplexa vaccination challenges responses parasites vaccines Parasitology
Divisions:
Page Range: pp. 392-398
Journal or Publication Title: Iranian Journal of Parasitology
Journal Index: ISI
Volume: 13
Number: 3
ISSN: 1735-7020
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/48

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