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Subcloning and Expression of Leishmania major (MRHO/IR/75/ER) P4 Gene

Shaddel, Minoo and Oormazdi, Hormozed and Akhlaghi, Lame and Kazemi, Bahram and Bandehpour, Mojgan (2013) Subcloning and Expression of Leishmania major (MRHO/IR/75/ER) P4 Gene. Journal of Archives in Military Medicine, 1 (1). pp. 19-23. ISSN 2345-5071

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Abstract

Background: Leishmania major p4 gene is localized in the endoplasmic reticulum at the intracellular amastigote stage. Objectives: We expressed this gene for possible future vaccine preparation, drug target studies, and leishmaniasis serodiagnosis test. Materials and Methods: The Leishmania major (MRHO/IR/75/ER) p4 gene, which had been subcloned into the pQE-30 expression vector, was induced by IPTG. Recombinant protein was confirmed by SDS-PAGE followed by a double diffusion and western blot using an anti His-tag antibody or human antibody. Results: Production of protein with approximately 35 kDa molecular weight in E. coli M15 transformed cells was confirmed using SDS-PAGE, which reacted with antibodies of leishmaniasis serum using double diffusion and western blot tests and either for anti His-tag antibody using western blot. Conclusions: We have expressed the Iranian L. major p4 gene successfully and are ready to continue the research for vaccine production. Positive results from the western blot and double diffusion test suggest the hypothesis of using this Ag in diagnostic tests. We used the pQE-30 plasmid as an expression vector, which has high quantity of expressed p4 protein Keywords: p4 gene; Gene Expression; Leishmania major; Subcloning

Item Type: Article
Subjects: WA Public Health
Depositing User: ePrint Admin Admin
Date Deposited: 09 Oct 2016 11:53
Last Modified: 09 Oct 2016 11:53
URI: http://eprints.ajaums.ac.ir/id/eprint/3597

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