Heterologous Expression of Hemagglutinin-Neuraminidase Protein from Newcastle Disease Virus Strain AF2240 in Centella Asiatica

Abstract

We explored the use of the medicinally important plant Centella asiatica for expression of hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) strain AF2240. HN protein is the principal target for subunit vaccine development against NDV. The full-length HN gene was cloned into a plant expression construct driven by the CaMV 35S promoter and C-terminal fusion of green fluorescence protein (GFP) as reporter system. The recombinant expression construct was transformed via particle bombardment into C. asiatica callus. Transformants were screened using GFP and selected on MS medium supplemented with 15 mg/l hygromycin. The ~1.8 kb HN mRNA transcript was detected on the putative transformants using RT-PCR. The presence of HN protein expression was further confirmed through dot blot analysis using anti-NDV chicken serum. Here we report, for the first time, the use of a novel medicinal plant as a new platform for HN protein expression.K Ke ey y w wo or rd ds s: : Antigen expression, edible vaccine, hemagglutinin-neuraminidase, medicinal plant, Newcastle disease virus.

Early Stage Development of a Newcastle Disease Vaccine Candidate in Corn

“…Additionally, the F and HN proteins expressed in transgenic maize plants were detected and quantified using anti-NDV antisera in ELSIA (38). The levels of expression of F and HN proteins in both leaves and seeds were in accordance with protein levels reported by other researchers (23,42). The orally fed, plant-derived NDV protein was able to induce a specific immune response in chickens compared to injected E. coli-expressed F and HN proteins and a commercially available NDV vaccine.…”

“…The HN protein plays a significant role in infection by facilitating virus attachment to host cells via sialic acid receptors. These proteins regulate the virulence of NDV and are considered immunogens for the development of any potent vaccine against NDV (23).…”

Comparison of Two Set Pathotypic-Specific Primers to Detect Newcastle Disease Virus

“…Determining the pathotype of NDV is generally carried out by isolating the virus from embryonated chicken eggs (ECEs), then testing it on Species Pathogen Free (SPF) chickens to measure the virulence of the virus using the intracerebral pathogenicity index (ICPI), an intravenous pathotype index (IVPI), and mean time to death (MDT) (OIE, 2012;Cattoli et al, 2011). Molecular techniques such as Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) have been developed by (Farooq et al, 2014;Lai et al, 2012;Rabalski et al, 2014) to identify the NDV. Amino acid sequencing is a further step that must be carried out to determine the pathotype of the NDV molecularly (Xiao et al,.…”