Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice
The current production of the Japanese encephalitis virus (JEV) vaccine is based on animal cells, where various risk factors for human health should be resolved. This study used a transient expression system to express the chimeric protein composed of antigenic epitopes from the JEV envelope (E) pro...
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MDPI AG
2023-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/14/11643 |
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| author | Jae-Wan Jung Pil-Gu Park Won-Kyung Lee Jun-Hye Shin Mi-Hwa Jang Eun-Hye Seo Timothy An Young Beom Kim Myeong Hee Moon Seuk-Keun Choi Jee Sun Yun Kee-Jong Hong Seong-Ryong Kim |
| author_facet | Jae-Wan Jung Pil-Gu Park Won-Kyung Lee Jun-Hye Shin Mi-Hwa Jang Eun-Hye Seo Timothy An Young Beom Kim Myeong Hee Moon Seuk-Keun Choi Jee Sun Yun Kee-Jong Hong Seong-Ryong Kim |
| author_sort | Jae-Wan Jung |
| collection | DOAJ |
| description | The current production of the Japanese encephalitis virus (JEV) vaccine is based on animal cells, where various risk factors for human health should be resolved. This study used a transient expression system to express the chimeric protein composed of antigenic epitopes from the JEV envelope (E) protein in <i>Nicotiana benthamiana</i>. JEV multi-epitope peptide (MEP) sequences fused with FLAG-tag or 6× His-tag at the C- or N-terminus for the purification were introduced into plant expression vectors and used for transient expression. Among the constructs, vector pSK480, which expresses MEP fused with a FLAG-tag at the C-terminus, showed the highest level of expression and yield in purification. Optimization of transient expression procedures further improved the target protein yield. The purified MEP protein was applied to an ICR mouse and successfully induced an antibody against JEV, which demonstrates the potential of the plant-produced JEV MEP as an alternative vaccine candidate. |
| first_indexed | 2024-03-11T00:59:21Z |
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| id | doaj.art-101292bbc59c42229249daab6d44775a |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T00:59:21Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-101292bbc59c42229249daab6d44775a2023-11-18T19:42:42ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-07-0124141164310.3390/ijms241411643Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in MiceJae-Wan Jung0Pil-Gu Park1Won-Kyung Lee2Jun-Hye Shin3Mi-Hwa Jang4Eun-Hye Seo5Timothy An6Young Beom Kim7Myeong Hee Moon8Seuk-Keun Choi9Jee Sun Yun10Kee-Jong Hong11Seong-Ryong Kim12Department of Life Science, Sogang University, Seoul 04107, Republic of KoreaDepartment of Microbiology, Gachon University College of Medicine, Incheon 21936, Republic of KoreaDepartment of Life Science, Sogang University, Seoul 04107, Republic of KoreaDepartment of Life Science, Sogang University, Seoul 04107, Republic of KoreaDepartment of Life Science, Sogang University, Seoul 04107, Republic of KoreaDepartment of Microbiology, Gachon University College of Medicine, Incheon 21936, Republic of KoreaDepartment of Microbiology, Gachon University College of Medicine, Incheon 21936, Republic of KoreaDepartment of Chemistry, Yonsei University, Seoul 03722, Republic of KoreaDepartment of Chemistry, Yonsei University, Seoul 03722, Republic of KoreaEubiologics Co., Seoul 06026, Republic of KoreaEubiologics Co., Seoul 06026, Republic of KoreaDepartment of Microbiology, Gachon University College of Medicine, Incheon 21936, Republic of KoreaDepartment of Life Science, Sogang University, Seoul 04107, Republic of KoreaThe current production of the Japanese encephalitis virus (JEV) vaccine is based on animal cells, where various risk factors for human health should be resolved. This study used a transient expression system to express the chimeric protein composed of antigenic epitopes from the JEV envelope (E) protein in <i>Nicotiana benthamiana</i>. JEV multi-epitope peptide (MEP) sequences fused with FLAG-tag or 6× His-tag at the C- or N-terminus for the purification were introduced into plant expression vectors and used for transient expression. Among the constructs, vector pSK480, which expresses MEP fused with a FLAG-tag at the C-terminus, showed the highest level of expression and yield in purification. Optimization of transient expression procedures further improved the target protein yield. The purified MEP protein was applied to an ICR mouse and successfully induced an antibody against JEV, which demonstrates the potential of the plant-produced JEV MEP as an alternative vaccine candidate.https://www.mdpi.com/1422-0067/24/14/11643Japanese encephalitis virus (JEV)multi-epitope peptide (MEP)recombinant proteintransient expression<i>Nicotiana benthamiana</i> |
| spellingShingle | Jae-Wan Jung Pil-Gu Park Won-Kyung Lee Jun-Hye Shin Mi-Hwa Jang Eun-Hye Seo Timothy An Young Beom Kim Myeong Hee Moon Seuk-Keun Choi Jee Sun Yun Kee-Jong Hong Seong-Ryong Kim Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice International Journal of Molecular Sciences Japanese encephalitis virus (JEV) multi-epitope peptide (MEP) recombinant protein transient expression <i>Nicotiana benthamiana</i> |
| title | Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice |
| title_full | Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice |
| title_fullStr | Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice |
| title_full_unstemmed | Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice |
| title_short | Production of Plant-Derived Japanese Encephalitis Virus Multi-Epitope Peptide in <i>Nicotiana benthamiana</i> and Immunological Response in Mice |
| title_sort | production of plant derived japanese encephalitis virus multi epitope peptide in i nicotiana benthamiana i and immunological response in mice |
| topic | Japanese encephalitis virus (JEV) multi-epitope peptide (MEP) recombinant protein transient expression <i>Nicotiana benthamiana</i> |
| url | https://www.mdpi.com/1422-0067/24/14/11643 |
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