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The role of GlcNAc-PI-de-N-acetylase gene by gene knockout through homologous recombination and its consequences on survival, growth and infectivity of Leishmania major in in vitro and in vivo conditions

Almani, P.G.N. and Sharifi, I. and Kazemi, B. and Babaei, Z. and Bandehpour, M. and Salari, S. and Dezaki, E.S. and Tohidi, F. and Mohammadi, M.A. (2016) The role of GlcNAc-PI-de-N-acetylase gene by gene knockout through homologous recombination and its consequences on survival, growth and infectivity of Leishmania major in in vitro and in vivo conditions. Acta Tropica, 154. pp. 63-72.

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Abstract

At present, there are no efficacious vaccines or effective drugs against leishmaniasis; therefore new and innovative control methods are urgently required. One way to achieve this important goal is through using reverse genetic engineering to evaluate important enzymes, proteins and macromolecules. One of the most important enzymes for Glycosylphosphatidylinositol (GPI) biosynthetic pathways is GlcNAc-PI-de-N-acetylase (GPI12). The molecular constructs were cloned in Escherichia coli strain Top 10 and confirmed by molecular methods and were transfected by electroporation into Leishmania major. We demonstrated that two alleles of the GPI12 gene in L. major were successfully removed and enabling the generation of a null mutant, which supports the idea that GPI12 is not an essential gene for the growth and survival of Leishmania and the homozygous knockouts of Leishmania are able to survive. We were able to produce a mutant parasite that caused no damaged to the host. Further investigations are essential to check the safety profile in laboratory animals. © 2015 Elsevier B.V.

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: enzyme; genetic analysis; genetic engineering; growth; infectivity; leishmaniasis; molecular analysis; protein; survival, allele; animal cell; animal experiment; animal model; animal tissue; Article; bacterial strain; controlled study; Escherichia coli; female; gene inactivation; genetic engineering; GPI12 gene; growth; homologous recombination; in vitro study; in vivo study; Leishmania major; leishmaniasis; mouse; mutant; nonhuman; parasite survival, Animalia; Escherichia coli; Leishmania major
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 08 Sep 2016 05:00
Last Modified: 13 Sep 2016 07:45
URI: http://eprints.goums.ac.ir/id/eprint/4589

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