Heterologous Expression of Human IL-29 (IFN-λ1) in Iranian Lizard Leishmania
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Abstract:
Background: Interferons with diffrent functions such as antiviral, antiproliferative and immunomodulatory actions are effctive medications for a number of diseases. One of these new interferons (IFNs) is Interleukin-29 (IL-29) belongs to the family of IFN-λ has antiviral activity and its potent in accompanying with IFN-α in treatment of HCV infection has been evaluated (clinical trial phase II). Recombinant IL-29 has been previously produced in multiple expression systems but in this study we cloned and expressed this protein in an Iranian Lizard leishmania (I.L.L) for the fist time. Objectives: The Main objective of this research was to evaluate expression of functional human IL-29 in domestic Lizard leishmania as an alternative eukaryotic expression system. Materials and Methods: The IL-29 expression cassette was constructed into a pLEXSY vector. The leishmania cells were transfected by electroporation. After selection of transfectants, the protein expression was evaluated at RNA and protein levels. Results: Expression cassette was successfully transfected to leishmania cells and expression of recombinant IL-29 was proved by RT-PCR and western blotting. Purifid protein showed 20% activity compared to standard protein. Enzymatic removal of N-glycan resulted to the shift of protein mobility on SDS-PAGE. Conclusions: Easy handling and culture of this eukaryotic host and mammalian cell like posttranslational modifiations are the main advantages of this expression host, but the expression yield of this protein is very low and it seems to be not economic for large-scale production.
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Journal title
volume 11 issue 3
pages 168- 174
publication date 2013-07-01
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