https://scholars.lib.ntu.edu.tw/handle/123456789/630240
標題: | Genetic transformation and expression of hemagglutinin gene from avian influenza virus in carrot (Daucus carota) | 作者: | Su, Yi Chen Huang, Pung Ling YI-YIN DO |
關鍵字: | Agrobacterium-mediated transformation | Codon-optimization | Plant-based oral vaccine | Production of recombinant protein | 公開日期: | 1-一月-2023 | 來源出版物: | Journal of Plant Biochemistry and Biotechnology | 摘要: | Avian influenza is a disease of poultry that causes economic losses to poultry farms. Conventional vaccine production requires chicken embryos for virus proliferation with time-consuming and also affected by egg supply during the avian influenza virus outbreak. There are several advantages of plant cells as bio-factories for vaccine production, including low cost, low risk of contamination, and even edible. In this study, we chose carrot (Daucus carota) as a bioreactor to produce hemagglutinin (HA), an avian influenza virus antigen protein. HA gene linked with heat-labile enterotoxin B subunit (LTB) gene whose product plays the role of adjuvant protein was codon optimized with codon usage bias of carrot for high-level expression. The native and codon-optimized LTB–HA were transformed into carrots by Agrobacterium-mediated transformation. T-DNA insertion of two version constructs was confirmed by polymerase chain reaction. Integration of codon-optimized LTB–HA gene in all transgenic carrot lines was identified as a single copy by Southern blot analysis. LTB–HA protein expressed in carrot cells was determined through enzyme-linked immunoassay. Codon-optimized LTB–HA gene showed a better expression level than the native LTB–HA gene in transgenic carrots. The highest expression level of LTB–HA in codon-optimized LTB–HA-transformed carrot taproot reached up to 0.0108% of total soluble protein. This study is the first report about expression of HA antigen of H5N2 subtype AIV in a horticultural crop. Our transgenic carrots have successfully produced HA protein which shows great potential to become an edible vaccine against avian influenza virus. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/630240 | ISSN: | 09717811 | DOI: | 10.1007/s13562-023-00840-6 |
顯示於: | 園藝暨景觀學系 |
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