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J Vet Med Sci. 2018 Sep 11. doi: 10.1292/jvms.18-0291. [Epub ahead of print]
System for the heterologous expression of NS1 protein of H9N2 avian influenza virus in the ciliate Tetrahymena thermophila.
Zhu C1,2,3, Liu X4, Chi H4, Chen C1, Chen Z1, Fu G1, Gong H4, Huang Y1,3.
Author information
Abstract
Tetrahymena is commonly used as an alternative eukaryotic system for efficiently expressing heterologous genes. In this study, we inserted the non-structural (NS) 1 gene of avian influenza virus (AIV) into the shuttle vector pD5H8 and transformed conjugating T. thermophila with the recombinant plasmid pD5H8-NS1 by particle bombardment. Positive transformants were selected with paromomycin. We demonstrated that the NS1 protein could be expressed steadily following induction with cadmium in this Tetrahymena system. An enzyme-linked immunosorbent assay detection method was preliminary established using the expressed protein as coating antigens for serodiagnosis. This is the first study in which a Tetrahymena expression system was employed for the expression of the AIV NS1 protein, and it provides a good basis for the development of differential diagnostic kits and vaccines for the prevention and control of avian influenza.
KEYWORDS:
NS1 gene; Tetrahymena; avian influenza; heterologous expression; virus
PMID: 30210069 DOI: 10.1292/jvms.18-0291
System for the heterologous expression of NS1 protein of H9N2 avian influenza virus in the ciliate Tetrahymena thermophila.
Zhu C1,2,3, Liu X4, Chi H4, Chen C1, Chen Z1, Fu G1, Gong H4, Huang Y1,3.
Author information
Abstract
Tetrahymena is commonly used as an alternative eukaryotic system for efficiently expressing heterologous genes. In this study, we inserted the non-structural (NS) 1 gene of avian influenza virus (AIV) into the shuttle vector pD5H8 and transformed conjugating T. thermophila with the recombinant plasmid pD5H8-NS1 by particle bombardment. Positive transformants were selected with paromomycin. We demonstrated that the NS1 protein could be expressed steadily following induction with cadmium in this Tetrahymena system. An enzyme-linked immunosorbent assay detection method was preliminary established using the expressed protein as coating antigens for serodiagnosis. This is the first study in which a Tetrahymena expression system was employed for the expression of the AIV NS1 protein, and it provides a good basis for the development of differential diagnostic kits and vaccines for the prevention and control of avian influenza.
KEYWORDS:
NS1 gene; Tetrahymena; avian influenza; heterologous expression; virus
PMID: 30210069 DOI: 10.1292/jvms.18-0291