Middle East respiratory syndrome coronavirus (MERS-CoV) first emerged in 2012, and over 2000 infections and 800 deaths have been confirmed in 27 countries. However, to date, no commercial vaccine is available. In this study, structural proteins of MERS-CoV were expressed in silkworm larvae and Bm5 cells for the development of vaccine candidates against MERS-CoV and diagnostic methods. The spike (S) protein of MERS-CoV lacking its transmembrane and cytoplasmic domains (SΔTM) was secreted into the hemolymph of silkworm larvae using a bombyxin signal peptide and purified using affinity chromatography. The purified SΔTM forms small nanoparticles as well as the full-length S protein and has the ability to bind human dipeptidyl peptidase 4 (DPP4), which is a receptor of MERS-CoV. These results indicate that bioactive SΔTM was expressed in silkworm larvae. To produce MERS-CoV-like particles (MERS-CoV-LPs), the coexpression of spike proteins was performed in Bm5 cells and envelope (E) and membrane (M) proteins secreted E and M proteins extracellularly, suggesting that MERS-CoV-LPs may be formed. However, this S protein was not displayed on virus-like particles (VLPs) even though E and M proteins were secreted into the culture supernatant. By surfactant treatment and mechanical extrusion using S protein-or three structural protein-expressing Bm5 cells, S protein-displaying nanovesicles with diameters of approximately 100-200 nm were prepared and confirmed by immuno-TEM. The mechanical extrusion method is favorable for obtaining uniform recombinant protein-displaying nanovesicles from cultured cells. The purified SΔTM from silkworm larvae and S protein-displaying nanovesicles from Bm5 cells may lead to the development of nanoparticle-based vaccines against MERS-CoV and the diagnostic detection of MERS-CoV. DDP4, to inhibit the infection of cells by MERS-CoV (Corti et al., 2015;Jiang et al., 2014). In addition, the S protein and its receptor-binding domain are regarded as promising targets for the development of vaccines against MERS-CoV, even though no vaccine against MERS-CoV is yet commercially available (Ma et al., 2014a, b). The M protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) forms enveloped virus-like particles (VLPs) when

A new method of label free sensing approach with superior selectivity and sensitivity towards virlabel-freeon is presented here, employing the localized surface plasmon resonance (LSPR) behavior of gold nanoparticles (AuNPs) and fluorescent CdSeTeS quantum dots (QDs). Inorganic quaternary alloyed CdSeTeS QDs were capped with -cysteine via a ligand exchange reaction. Alternatively, citrate stabilized AuNPs were functionalized with 11-mercaptoundecanoic acid to generate carboxylic group on the gold surface. The carboxylic group on the AuNPs was subjected to bind covalently with the amine group of-cysteine capped CdSeTeS QDs to form CdSeTeS QDs/AuNPs nanocomposites. The fluorescence of CdSeTeS QDs/AuNPs nanocomposite shows quenched spectrum of CdSeTeS QDs at 640 nm due to the close interaction with AuNPs. However, after successive addition of norovirus-like particles (NoV-LPs), steric hindrance-induced LSPR signal from the adjacent AuNPs triggered the fluorescence enhancement of QDs in proportion to the concentration of the target NoV-LPs. A linear range of 10 to 10 g mL NoV-LPs with a detection limit of 12.1 × 10 g mL was obtained. This method was further applied on clinically isolated norovirus detection, in the range of 10-10 copies mL with a detection limit of 95.0 copies mL, which is 100-fold higher than commercial ELISA kit. The superiority of the proposed sensor over other conventional sensors is found in its ultrasensitive detectability at low virus concentration even in clinically isolated samples. This proposed detection method can pave an avenue for the development of high performance and robust sensing probes for detection of virus in biomedical applications.

Nanobiomaterials such as enveloped virus-like particles with specificity can serve a dual function of vaccination and drug delivery system. Here, we expressed colon carcinoma cell-targeting chimeric virus-like particles (VLPs) made using two capsid proteins, gag and M1 from influenza virus A/swine flu/Iowa/15/30/H1N1 in silkworms. These chimeric VLPs displayed a glycosylphosphatidylinositol-anchored single-chain variable fragment region targeting colon carcinoma cells, and their shape was smooth, with an average particle size of 21 nm in diameter. Large unilamellar vesicles made from DOPC:DOPA (2:1) containing calcein-AM (10 μM) or doxorubicin (13.7 nM) were used to package chimeric VLPs. VLPs showed high specificity in targeting cancer cells and delivered the dye and drug to cells successfully. Chimeric VLPs were injected into BALB/c mice, and the serum showed specificity for M1 protein as a model.

16Rous sarcoma virus group antigen protein based virus like particles (VLPs)

To examine the effect of 2-azahypoxanthine (AHX) on rice plant (Oryza sativa L. cv. Nipponbare) growth, we carried out pot and field experiments. AHX was applied at 50 μM for two weeks at four growth stages (transplanting, tillering, panicle formation and ripening stages) in the pot experiment, and 1 mM AHX at three stages [at rising of seedling in nursery boxes (seedling treatment), transplanting and panicle formation stages] in the field experiment. Both pot and field experiments showed a tendency toward increased panicle number (PN), culm length (CL) and plant dry weights with AHX treatments. Brown rice yields were also improved by AHX treatments, especially when applying at stages of tillering and panicle formation and seedling and transplanting during pot and field experiments, respectively. In the latter, yield increased drastically up to 18.7, 15.8, 9.6 and 5.8% of control. However, panicle length and 1000-grain weight were not affected by AHX application. These results suggested that AHX increased the brown rice yield through its effects on PN and/or CL.