Baculovirus-mediated gene expression in chicken primary cells

Avian Diseases
Wenxiang PingZhuangwei Lou

Abstract

A recombinant baculovirus was constructed containing an expression cassette with a reporter gene, green fluorescent protein, directed by a constitutive mammalian promoter: a human cytomegalovirus immediate early promoter/enhancer (CMV-IE). High titer virus was prepared with ultracentrifugation. Efficient gene delivery and expression were observed in the virus-treated chicken primary culture, myoblast cells, and whole embryonic fibroblast cells. It was noticed that an addition of sodium butyrate (a selective histone deacetylase inhibitor) to viral transduction medium extremely enhanced the reporter-gene expression. However, there is no effect of presence of trichostatin A observed. To maximize the reporter-gene expression, the baculoviral infection condition was optimized with both cell types. Our approaches demonstrated that recombinant baculovirus could efficiently deliver its genome DNA into chicken primary cells and that CMV-IE, a mammalian-cell-active promoter, was functional in chicken primary cells and could direct a high level of gene expression. Clearly, the recombinant baculovirus provides an alternative means for foreign gene delivery into avian cells.

References

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Citations

Mar 22, 2007·Molecular Therapy : the Journal of the American Society of Gene Therapy·Ding-Gang YangYu-Chen Hu
Apr 29, 2015·Viruses·Kaisa-Emilia MakkonenSeppo Ylä-Herttulala
Dec 4, 2012·Fish & Shellfish Immunology·Renyu XueChengliang Gong
Mar 1, 2011·Journal of Virological Methods·Fengtao HuangXueqin Liu
Apr 18, 2016·Molecular Biology Reports·Bo LiuChengliang Gong
Nov 30, 2018·International Journal of Molecular Sciences·Qian WangTiansheng Chen
Jun 1, 2017·Journal, Genetic Engineering & Biotechnology·Mohamed A El-MogyYousef Haj-Ahmad

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Methods Mentioned

BETA
PCR
transfection

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