Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i>
The coat protein (CP) is an important structural protein that plays many functional roles during the viral cycle. In this study, the CP of pepper mild mottle virus (PMMoV) was genetically fused to GFP using the foot-and-mouth disease virus peptide 2A linker peptide and the construct (PMMoV-GFP<su...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-03-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/13/7/928 |
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| author | Mengting Jiao Yueyan Yin Yanzhen Tian Jianing Lei Lin Lin Jian Wu Yuwen Lu Hongying Zheng Fei Yan Jianguang Wang Jiejun Peng |
| author_facet | Mengting Jiao Yueyan Yin Yanzhen Tian Jianing Lei Lin Lin Jian Wu Yuwen Lu Hongying Zheng Fei Yan Jianguang Wang Jiejun Peng |
| author_sort | Mengting Jiao |
| collection | DOAJ |
| description | The coat protein (CP) is an important structural protein that plays many functional roles during the viral cycle. In this study, the CP of pepper mild mottle virus (PMMoV) was genetically fused to GFP using the foot-and-mouth disease virus peptide 2A linker peptide and the construct (PMMoV-GFP<sup>2A</sup>) was shown to be infectious. The systemic spread of the virus was monitored by its fluorescence in infected plants. Electron microscopy and immunocolloidal gold labelling confirmed that PMMoV-GFP<sup>2A</sup> forms rod-shaped particles on which GFP is displayed. Studies of tissue ultrastructure and virion self-assembly confirmed that PMMoV-GFP<sup>2A</sup> could be used to monitor the real-time dynamic changes of CP location during virus infection. Aggregations of GFP-tagged virions appeared as fluorescent plaques in confocal laser microscopy. Altogether, PMMoV-GFP<sup>2A</sup> is a useful tool for studying the spatial and temporal changes of PMMoV CP during viral infection. |
| first_indexed | 2024-04-24T10:36:48Z |
| format | Article |
| id | doaj.art-6316d56703d14eeaa514f08d76ff32f8 |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-04-24T10:36:48Z |
| publishDate | 2024-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-6316d56703d14eeaa514f08d76ff32f82024-04-12T13:24:38ZengMDPI AGPlants2223-77472024-03-0113792810.3390/plants13070928Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i>Mengting Jiao0Yueyan Yin1Yanzhen Tian2Jianing Lei3Lin Lin4Jian Wu5Yuwen Lu6Hongying Zheng7Fei Yan8Jianguang Wang9Jiejun Peng10State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaInstitute of Biotechnology and Germplasm Resources, Yunnan Academy of Agricultural Sciences, Kunming 650223, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaBiocontrol Engineering Research Center of Crop Disease & Pest of Yunnan Province, School of Ecology and Environmental Science, Yunnan University, Kunming 650091, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agroproducts, Institute of Plant Virology, Ningbo University, Ningbo 315211, ChinaThe coat protein (CP) is an important structural protein that plays many functional roles during the viral cycle. In this study, the CP of pepper mild mottle virus (PMMoV) was genetically fused to GFP using the foot-and-mouth disease virus peptide 2A linker peptide and the construct (PMMoV-GFP<sup>2A</sup>) was shown to be infectious. The systemic spread of the virus was monitored by its fluorescence in infected plants. Electron microscopy and immunocolloidal gold labelling confirmed that PMMoV-GFP<sup>2A</sup> forms rod-shaped particles on which GFP is displayed. Studies of tissue ultrastructure and virion self-assembly confirmed that PMMoV-GFP<sup>2A</sup> could be used to monitor the real-time dynamic changes of CP location during virus infection. Aggregations of GFP-tagged virions appeared as fluorescent plaques in confocal laser microscopy. Altogether, PMMoV-GFP<sup>2A</sup> is a useful tool for studying the spatial and temporal changes of PMMoV CP during viral infection.https://www.mdpi.com/2223-7747/13/7/9282Apepper mild mottle viruscoat proteinself-assembly |
| spellingShingle | Mengting Jiao Yueyan Yin Yanzhen Tian Jianing Lei Lin Lin Jian Wu Yuwen Lu Hongying Zheng Fei Yan Jianguang Wang Jiejun Peng Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i> Plants 2A pepper mild mottle virus coat protein self-assembly |
| title | Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i> |
| title_full | Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i> |
| title_fullStr | Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i> |
| title_full_unstemmed | Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i> |
| title_short | Adoption of the 2A Ribosomal Skip Principle to Track Assembled Virions of Pepper Mild Mottle Virus in <i>Nicotiana benthamiana</i> |
| title_sort | adoption of the 2a ribosomal skip principle to track assembled virions of pepper mild mottle virus in i nicotiana benthamiana i |
| topic | 2A pepper mild mottle virus coat protein self-assembly |
| url | https://www.mdpi.com/2223-7747/13/7/928 |
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