Cell Surface Display of Red-Grouper Nervous Necrosis Virus Capsid Protein on &amp;lt;i&amp;gt;Pichia pastoris&amp;lt;/i&amp;gt;

Intamaso, Uraiwan; Chutoam, Palatip; Poomipak, Witthaya; Pirarat, Nopadon

doi:10.4236/aim.2018.811055

Cited by 3 publications

(10 citation statements)

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“…The generation of Pichia pastoris surface‐displayed viral capsid proteins was in hopes of developing an oral vaccine against red‐grouper‐nervous‐necrosis virus (RGNNV) in fish (Intamaso et al . 2018). Zhao et al (2017) designed an oral yeast vaccine which surface‐displayed the glycoprotein (G) of Chinese infectious hematopoietic necrosis virus (IHNV) strain and delivered orally to rainbow trout.…”

Section: Application In Aquaculturementioning

confidence: 99%

Surface display system for probiotics and its application in aquaculture

Yao

Yang

Gao

et al. 2020

Reviews in Aquaculture

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Aquaculture is a fast‐growing and rapidly expanding industry in the world. Probiotics are widely used in aquaculture and have provided benefits to aquatic animal health, and it is also a promising alternative to antibiotics for control of fish diseases. With the development of biotechnology, new expression systems and novel techniques for the surface display of heterologous proteins in surface of probiotics cells have been developed. This review provides an overview of the systems and strategies for displaying functional proteins on the surface of probiotics commonly used in aquaculture, which are Bacillus subtilis, Bacillus thuringiensis, lactic acid bacteria represented by Lactococcus lactis and yeast. Their applications in aquaculture especially for oral vaccine development afforded by this technology and prospects and challenges associated with this technology are also highlighted.

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Section: Application In Aquaculturementioning

confidence: 99%

Surface display system for probiotics and its application in aquaculture

Yao

Yang

Gao

et al. 2020

Reviews in Aquaculture

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show abstract

“…The RNA2 gene was cloned from RG-NNV and used to modify the P. pastoris strain GS115 (Mut + His -) genetically (Intamaso et al, 2018). The expression plasmid using heterologous protein expression was pPIC9K purchased from Invitrogen (Carlsbad, CA, USA).…”

Section: Strains Growth Medium and Plasmidmentioning

confidence: 99%

“…The recombinant P. pastoris GS115 (Mut s His + ) was generated by fusing the RG-NNV RNA2 gene to S. cerevisiae AGα1 gene coding for α-agglutinin (Intamaso et al, 2018). The recombinant and a negative control yeast containing only the AGα-1 gene were cultured on YEPD agar and incubated at 30 °C for 72 hr.…”

Section: Optimization Of Cell Growth and Rna2 In A Shake Flaskmentioning

confidence: 99%

“…This study was approved by the Ethics Committee of BUU-IBC Burapha University (Approval no.26/2558). Intamaso et al (2018) reported that RNA2 capsid proteins of RG-NNV linked to AGα-1 of S. cerevisiae were displayed at low levels on the P. pastoris cell surface. However, the current study demonstrated that growing yeast under optimized conditions successfully increased the levels of fusion proteins displayed on the yeast cell surface.…”

Section: Ethics Statementmentioning

confidence: 99%

“…However, in another study, the number of antigens detected was relatively low and likely inadequate to boost any immune response when applied in fish. Thus the current work determined the environmental factors affecting the production of RNA2/AGα-1 fusion proteins displayed on the P. pastoris cell surface (Intamaso et al, 2018). The approach obtained under shake-flask culture should facilitate further optimization of large-scale production of the oral vaccine against RG-NNV in fish.…”

Section: Introductionmentioning

confidence: 99%

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Optimized expression of capsid protein from red-grouper necrosis virus displayed on Pichia pastoris in shake flasks

2023

ANRES

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Importance of the work: High antigen levels on the yeast cell surface are critical in designing oral vaccines against viruses. Objectives: To investigate the environmental factors affecting the production of RNA2/ AGα-1, a displayed fusion protein. Materials & Methods:The recombinant Pichia pastoris was cultivated in shake-flask culture under varying conditions and induced with methanol for 168 hr. Total cell proteins were used as a measure of cell population growth. The expression of the RNA2 fusion protein was also characterized by western blot, immunofluorescence and enzyme-linked immunosorbent assay (ELISA). Results: The recombinant P. pastoris (Mut s ) strain produced optimal growth and product yields using 72 hr post-induction and 0.5% methanol. Prior to induction, the yeast relied on glycerol as the sole carbon source to increase biomass under favorable conditions: 23 °C in an Erlenmeyer flask (5 L) containing 1 L buffered minimal glycerol complex medium (pH 6.0) with vigorous shaking (300 revolutions per minute). Using these optimal conditions, western blot analysis revealed that the RNA2 fusion protein had a molecular weight of approximately 73 kDa. These proteins were localized on the yeast cell surface, as proven based on immunofluorescence labeling at a minimum of 1,000 ng and measured using ELISA. Main finding: The method developed successfully increased the expression of capsid proteins from the virus on the yeast cell surface in a shake flask culture. Large-scale production is required before investigating the potential use of the fusion protein as an oral vaccine in fish.

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First Report of Viral Nervous Necrosis in Asian Sea Bass, Lates Calcarifer Cultured in Sri Lanka

Fouzi,

Sakajamary

2023

J. Agro-Technol. Rural Sci.

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Viral Nervous Necrosis (VNN) caused by Betanodavirus is a devastating disease in aquatic animals. The virus infects both marine and freshwater fish worldwide. Vacuolating necrosis of neural cells of the brain, the retina of the eye, and the spinal cord of the infected fish are the primary histological lesions of the condition. It causes up to 100% mortality in larvae and juvenile fish and can cause significant death in adult fish. The present study detected viral nervous necrosis in larvae and fry of Asian sea bass (Lates calcarifer) with progressive mortality of up to 95% in one week during the Northeast monsoon when the mean water temperature was 27 to 29°C. Histopathological examination of the moribund fish revealed extensive vacuolation and gliosis in the olfactory bulb, the optic lobe of the forebrain, and the inner and outer layer of the retina. Furthermore, tissues of the brain and the retina had intracellular inclusion bodies suggesting viral etiology, further justified by the negative results of the bacterial and parasitic examinations. The Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) test additionally confirms the etiology diagnosis using specific primers designed previously. The histopathology and RT-PCR results suggest that the mortalities of Asian sea bass were due to the VNN. The present finding is the first report of the VNN associated with mass mortalities in Asian seabass cultured in Sri Lanka. These crucial findings emphasize the need for quarantine and control strategies to prevent the spread of the virus and outbreak of the disease.

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Cell Surface Display of Red-Grouper Nervous Necrosis Virus Capsid Protein on <i>Pichia pastoris</i>

Cited by 3 publications

References 35 publications

Surface display system for probiotics and its application in aquaculture

Surface display system for probiotics and its application in aquaculture

Optimized expression of capsid protein from red-grouper necrosis virus displayed on Pichia pastoris in shake flasks

First Report of Viral Nervous Necrosis in Asian Sea Bass, Lates Calcarifer Cultured in Sri Lanka

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