Construction of Single-walled Carbon Nanotube-loaded Koi Herpes Virus ORF149 Nucleic Acid Vaccine

doi:10.13560/j.cnki.biotech.bull.1985.2019-0688

Abstract

Abstract: In order to develop a new type of highly effective vaccine against Koi Herpesvirus（KHV）disease,a single-walled carbon nanotube（SWCNT）was used as a transport vector to construct a KHV ORF149 nucleic acid vaccine. The recombinant plasmid pcDNA3.1（+）-ORF149 was firstly constructed,and its expression was confirmed by transient transfection and immunoblot analysis. Then,the recombinant plasmid was conjugated to SWCNTs by 1,3-dipolar cycloaddition reaction. Scanning electron microscopy and nucleic acid electrophoresis was conducted to identify whether the conjugation was successful. The results showed that specific signals were detected with constructed recombinant plasmid in both immunoblot analysis and indirect immunofluorescence assay after transfection. In the nucleic acid agarose gel electrophoresis,the electrophoretic band of the constructed recombinant vaccine disappeared. Under the observation of scanning electron microscopy,the morphologies of the conjugated SWCNTs with the recombinant plasmid and blank SWCNTs were significantly different.

Key words: single-walled carbon nanotubes, KHV ORF149, DNA vaccine

HU Feng, WANG Qing, LI Ying-ying, ZENG Wei-wei, WANG Gao-xue, ZHU Bin, WANG Ying-ying, YIN Ji-yuan. Construction of Single-walled Carbon Nanotube-loaded Koi Herpes Virus ORF149 Nucleic Acid Vaccine[J]. Biotechnology Bulletin, 2020, 36(2): 206-213.

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