Genetic transformation and expression of hemagglutinin gene from avian influenza virus in carrot (Daucus carota)

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...

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Veröffentlicht in:Journal of plant biochemistry and biotechnology 2023-09, Vol.32 (3), p.550-559
Hauptverfasser: Su, Yi-Chen, Huang, Pung-Ling, Do, Yi-Yin
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Sprache:eng
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Zusammenfassung: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.
ISSN:0971-7811
0974-1275
DOI:10.1007/s13562-023-00840-6