Influenza hemagglutinin expression in Nicotiana tabacum and Nicotiana benthamiana.
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Chandler, Garvin Lee.
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Influenza infects millions of people each year. Hemagglutinin (HA) is the major surface protein of the influenza virus and the best target as a vaccine candidate. Transgenic plants have significant potential for proper expression of HA to serve as an influenza vaccine. An HA3 DNA construct for insertion into plants was developed using the binary vector, pE1802. Agrobacterium tumefaciens was transformed with the plasmid and both Nicotiana tabacum or Nicotiana benthamiana were transformed. To screen for HA3 expression, mRNA was isolated from first generation plants. Second generation plants were successfully screened for protein production using immunoblots to verify HA3 protein expression. A tobacco mosaic virus (TMV) vector (pJL36) was also produced expressing HA3. Recombinant constructs of the TMV vector were successfully screened using PCR. This work indicates the potential use of HA3-expressing TMV vector for transformation into A. tumefaciens, infection into N. benthamiana and the expression of gene products in these systems.