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Production of recombinant construction pBudCE4.1IFNβ-1a including recombinant Interferon beta gene and transfection into HEK293cell line

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Year: 2014
COI code: CIGS13_0239
Paper Language: English

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Authors Production of recombinant construction pBudCE4.1IFNβ-1a including recombinant Interferon beta gene and transfection into HEK293cell line

  Rahele Norouzi - Division of Genetics, Department of Biology, Faculty of Science, University of Isfahan, Isfahan, IR Iran
  Zohreh Hojati Najaf abadi - Division of Genetics, Department of Biology, Faculty of Science, University of Isfahan, Isfahan, IR Iran

Abstract:

Interferons(IFNs) are a group of secretory cytokines with a low molecular weight which possess broad range of biological effects, including antitumoral properties, adjust the immune system and regulate cell differentiation. Human IFNβ is afibroblastic glycoprotein produced in response to viral infections. In medicine, IFNβ used in order to autoimmune diseasestreatment such as multiple sclerosis, cell proliferation and angiogenesis inhibition. IFNβ has two analogs. Non-glycosylatedform, IFNβ-1b, and glycosylated form, IFNβ-1a. Object: According to the transient expression of IFNβ and its medical necessity, changes the gene in order to produce more and better quality protein, will be discussed. Method:In this study, the expression level of recombinant human IFNβ-1a gene including kozak conserved sequence and targeted mutations in 27 and 101 codons in gene coding region at both mRNA and protein levels will be examined. PBudCE4.1 is selected vector for this study which designed for the same expression of two genes in mammalian cell lines. HEK293 usedfor transfection of our designed vector. This selected cell line is known as one of the best choices for recombinant gene expression, producing variety of viral vectors and vaccines aims. In this examination, Real time PCR and protein diagnostic techniques such as Dot blot and Elisa will be considered. Conclusion: Low biological activity of IFNβ-1b produced in Ecoli and different physicochemical and immunogenicity properties compare to natural form, reveal the importance of protein glycosylation. In addition, mutations applied in order to increase stability, binding affinity and protein proper folding will be important

Keywords:

IFNβ, Multiple sclerosis, HEK293, pBudCE4.1, transfection

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COI code: CIGS13_0239

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Norouzi, Rahele & Zohreh Hojati Najaf abadi, 2014, Production of recombinant construction pBudCE4.1IFNβ-1a including recombinant Interferon beta gene and transfection into HEK293cell line, 12th Congress of Iranian Genetics Society, تهران, انجمن ژنتيك ايران, https://www.civilica.com/Paper-CIGS13-CIGS13_0239.htmlInside the text, wherever referred to or an achievement of this article is mentioned, after mentioning the article, inside the parental, the following specifications are written.
First Time: (Norouzi, Rahele & Zohreh Hojati Najaf abadi, 2014)
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Paper No.: 13235
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