Heterologous Expression of Human IL-29 (IFN-λ1)in Iranian Lizard Leishmania

被引:7
|
作者
Taromchi, Amir Hossein [1 ]
Kazemi, Bahram [1 ,2 ]
Mahmazi, Sanaz [3 ]
Bandehpour, Mojgan [1 ,2 ]
机构
[1] Shahid Beheshti Univ Med Sci, Dept Biotechnol, Tehran, Iran
[2] Shahid Beheshti Univ Med Sci, Cellular & Mol Biol Res Ctr, Tehran, Iran
[3] Islamic Azad Univ, Dept Genet, Zanjan Branch, Zanjan, Iran
关键词
Interferons; Interleukin-29; Recombinant; Leishmania; ANTITUMOR-ACTIVITY; INTERFERON-LAMBDA; PURIFICATION; TARENTOLAE; PROTEIN; SYSTEM;
D O I
10.5812/ijb.12468
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: Interferons with different functions such as antiviral, antiproliferative and immunomodulatory actions are effective 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 first 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. Purified 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 modifications 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. © 2013, National Institute of Genetic Engineering and Biotechnology; Licensee Kowsar Ltd.
引用
收藏
页码:168 / 174
页数:7
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