Volume 19, Issue 5 (8-2016)                   J Arak Uni Med Sci 2016, 19(5): 42-50 | Back to browse issues page

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Donyapoor F, Zeinoddini M, Saeedinia A R. Cloning and Expression of Recombinant Immunotoxin using Diphtheria Toxin and Granulocyte Colony Stimulating Factor (G-CSF). J Arak Uni Med Sci 2016; 19 (5) :42-50
URL: http://jams.arakmu.ac.ir/article-1-4210-en.html
1- Department of Sciences and Biotechnology, Malek- Ashtar University of Technology, Tehran, Iran.
2- PhD in Biochemistry, Department of Sciences and Biotechnology, Malek- Ashtar University of Technology, Tehran, Iran. , zeinoddini@modares.ac.ir
3- PhD in Genetics, Department of Sciences and Biotechnology, Malek- Ashtar University of Technology, Tehran, Iran.
Abstract:   (4846 Views)

Background: Immunotoxin (IT) is a directed toxin containing two distinct sections (immune and toxin parts) covalently bond using specific chemical or peptide linkers. The aim of this study is to produce a recombinant and hybrid protein containing diphtheria toxin (DT) and granulocyte colony stimulating factor (G-CSF).

Materials and Methods: According to the structure of the first commercial recombinant immunotoxin (Ontak, hybrid protein containing DT fused interlukine2), gene encoding of DT and G-CSF was amplified using specific primers and plasmid template of pET-IDZ3 (pET21 harboring the gene encoding ontak immunotoxine) and pET-GCSF (pET23 harboring G-CSF), respectively. The DT-GCSF fusion protein produced using soeing PCR and specific primers. Finally, pET-DT-GCSF construction prepared using subcloning of DT-GCSF into pET21a(+) and confirmed by sequencing, SDS-PAGE and western blot technique.

Results: Gene encoding of DT-GCSF inserted into NdeI/EcoRI site of pET21 and the construction of strain producing DT-GCSF recombinant immunotoxin was confirmed using customary methods.

Conclusion: The cytokine fusion protein, DT-GCSF, could be used for the inhibition of G-CSF receptor bearing cancer cells.

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Type of Study: Original Atricle | Subject: Basic Sciences
Received: 2016/01/24 | Accepted: 2016/06/11

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