Expression of an Avian Influenza Virus (H5N1) Hemagglutinin Gene in Transgenic Lotus corniculatus

Single‐Particle Cryo‐EM and 3D Reconstruction of Hybrid Nanoparticles with Electron‐Dense Components

“…After such normalization, each pixel value reflects signal‐to‐noise ratio (SNR). As shown in Figure , hybrid particles with simulated central “gold” of different intensities were generated and processed independently using jspr to evaluate their ability in giving correct initial models. As predicted, the interference of “gold” pixels with 2D alignment became more severe with the increase of “gold” pixel intensities; A “gold” core with intensity of 4 and higher above the icosahedral shell signal made the initial model building of phage T7 capsid II to fail (Figure ).…”

“…After these tests with simulated T7/“gold” particles, we summarized a strategy for single‐particle image processing of hybrid particles by combining maskGold.py masking with our truly independent single‐particle 3D reconstruction method . As illustrated in Figure , after preprocessing (i.e., particle picking and CTF parameters determination and correction), hybrid particles were first masked using maskGold.py resulting in two sets of particles, the normalized ones ( .norm , with the non‐gold pixels normalized to mean = 0 and sigma = 1) and the masked ones ( .masked ) derived from the normalized images ( .norm ) yet with gold pixels masked.…”

Production of vaccines for treatment of infectious diseases by transgenic plants

“…Highly pathogenic avian influenza is a deadly disease that had initially been affecting only poultry, thus causing great economic losses, but has also been representing a serious risk for human health since the first outbreak of highly pathogenic influenza (H5N1) in humans in Hong Kong in 1997 (Guo et al, 2012) in Asian countries as well as in Europe and Africa. Development of safe, cost-effective and efficacious vaccines is thus needed to reduce the economic impact of the disease on agriculture and prevent possible future pandemic outbreaks.…”

“…The main targets for subunit vaccine development have been its principal surface antigen hemagglutinin (HA), which plays a major role in functional activity of the virus and is also a main target for neutralizing antibodies of the host, and the M2e viral peptide, the extracellular domain of the viral protein M2, which is highly conserved between various influenza strains and therefore a promising candidate for a universal influenza vaccine. H5N1 HA has been expressed in N. benthamiana in full-length (Kalthoff et al, 2010) and in the form of VLPs (Landry et al, 2010), in Lotus corniculatus (Guo et al, 2012) and most recently, duckweed (Bertran et al, 2015). In addition, recombinant HA from A/Bar-headed Goose/Qinghai/1A/05 (clade 2.2) and A/Anhui/1/2005 (clade 2.3) have been expressed in N. benthamiana (Shoji et al, 2009).…”

“…34 % of them developed transient IgG and in some cases IgE against plant glyco-epitops but no hypersensitivity or allergy was observed. Guo et al (2012) were the first to successfully express biologically functional H5N1 HA in L. corniculatus, the gene expression being driven by CaMV 35S promoter, using Agrobacteriummediated transformation. Transformation frequency in successfully transformed cotyledon fragments was 58.8 % as determined by PCR or Southern blot analysis, while transformation of hypocotyl gave no results.…”