Construction of lentivirus-mediated stable expression of duck interferon-γ cell line

doi:10.16178/j.issn.0528-9017.20218494

Abstract

Abstract:

Interferon-γ is one of the cytokines with antiviral activity and immunomodulatory ability. To construct a cell line stably expressing duck interferon-γ, the gene of duck interferon gamma (dIFN-γ) was optimized referring to mouse codon and synthesized. Synthesed dIFN-γ was cloned into lentiviral expressing vector plenti-GIII-CMV-GFP-2A-Puro to construct recombinant plasmid plenti-IFN. The recombinant lentiviral rlenti-IFN was rescued by co-transfected plenti-IFN and helper plasmid into packaging cell 293T. After infection Chinese hamster ovary(CHO)cells with recombiant rlenti-IFN, 30 monoclonal cell lines with purinomycin resistance were selected by limited dilution and pressure screening. At last, one monoclonal cell line with the highest duIFN-γ mRNA level was selected by RT-qPCR method. One stable cell line was finally obtained by expanding culture. Western blot analysis indicated that duIFN-γ was stably expressed in this cell line. This study lays a foundation for studying duIFN-γ biological function, establishing duIFN-γ detection method and producing cheap interferon for duck or poultry.

Key words: duck interferon-γ, lentivirus, CHO cell, stable expression

CLC Number:

S858.32

CHEN Liu, NI Zheng, LIU Keshu, YE Weicheng, HUA Jionggang, YUN Tao, ZHU Yinchu, ZHANG Cun. Construction of lentivirus-mediated stable expression of duck interferon-γ cell line[J]. Journal of Zhejiang Agricultural Sciences, 2024, 65(1): 208-212.

Figures/Tables 3

References 21

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