System for the heterologous expression of NS1 protein of H9N2 avian influenza virus in the ciliate Tetrahymena thermophila

• Published in: Journal of Veterinary Medical Science Vol. 80; no. 10; pp. 1610 - 1618
• Main Authors: ZHU, Chunhua, LIU, Xiaodong, CHI, Hongshu, CHEN, Cuiteng, CHEN, Zhen, FU, Guanghua, GONG, Hui, HUANG, Yu
• Format: Journal Article
• Language: English
• Published: Japan JAPANESE SOCIETY OF VETERINARY SCIENCE 2018; Japan Science and Technology Agency; The Japanese Society of Veterinary Science
• Subjects: Antigens; Avian flu; avian influenza; Cadmium; Cloning, Molecular; Codon; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Gene Amplification; Gene Expression - drug effects; Genetic Vectors; heterologous expression; Influenza; Influenza A Virus, H9N2 Subtype - genetics; Microorganisms, Genetically-Modified; NS1 gene; NS1 protein; Paromomycin; Particle bombardment; Proteins; Protozoa; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Tetrahymena; Tetrahymena thermophila - genetics; Viral Nonstructural Proteins - genetics; Virology; virus; NS1 gene; heterologous expression; Tetrahymena; avian influenza; virus
• ISSN: 0916-7250; 1347-7439; 1347-7439
• DOI: 10.1292/jvms.18-0291

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.