Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector
A recombinant plasmid, pTXS.TH, was constructed to express the gene-encoding wasabi (Wasabia japonica) defensin with the potato virus X (PVX) vector. pTXS.TH allows the expression of defensin in the host Nicotiana benthamiana, and the defensin protein WT1 can be purified from virus-infected leaves b...
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| Format: | Article |
| Language: | English |
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The American Phytopathological Society
2001-02-01
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| Series: | Molecular Plant-Microbe Interactions |
| Subjects: | |
| Online Access: | https://apsjournals.apsnet.org/doi/10.1094/MPMI.2001.14.2.111 |
| _version_ | 1818647782338068480 |
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| author | Hiromasa Saitoh Akinori Kiba Masahiro Nishihara Saburo Yamamura Kazumi Suzuki Ryohei Terauchi |
| author_facet | Hiromasa Saitoh Akinori Kiba Masahiro Nishihara Saburo Yamamura Kazumi Suzuki Ryohei Terauchi |
| author_sort | Hiromasa Saitoh |
| collection | DOAJ |
| description | A recombinant plasmid, pTXS.TH, was constructed to express the gene-encoding wasabi (Wasabia japonica) defensin with the potato virus X (PVX) vector. pTXS.TH allows the expression of defensin in the host Nicotiana benthamiana, and the defensin protein WT1 can be purified from virus-infected leaves by heat treatment and affinity chromatography. WT1 exhibits strong antifungal activity toward the phytopathogenic fungi Magnaporthe grisea(50% inhibitory concentration [IC50] = 5 μg/ml) and Botrytis cinerea (IC50 = 20 μg/ml) but is weakly active against the phytopathogenic bacterium Pseudomonas cichorii. This virus-mediated expression system is a rapid and efficient method to produce and characterize antimicrobial proteins in plants. It is particularly useful for the study of proteins that are difficult to produce with other expression systems. |
| first_indexed | 2024-12-17T01:08:00Z |
| format | Article |
| id | doaj.art-7a5894f982674a9cb5f2b0884c28310e |
| institution | Directory Open Access Journal |
| issn | 0894-0282 1943-7706 |
| language | English |
| last_indexed | 2024-12-17T01:08:00Z |
| publishDate | 2001-02-01 |
| publisher | The American Phytopathological Society |
| record_format | Article |
| series | Molecular Plant-Microbe Interactions |
| spelling | doaj.art-7a5894f982674a9cb5f2b0884c28310e2022-12-21T22:09:13ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77062001-02-0114211111510.1094/MPMI.2001.14.2.111Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X VectorHiromasa SaitohAkinori KibaMasahiro NishiharaSaburo YamamuraKazumi SuzukiRyohei TerauchiA recombinant plasmid, pTXS.TH, was constructed to express the gene-encoding wasabi (Wasabia japonica) defensin with the potato virus X (PVX) vector. pTXS.TH allows the expression of defensin in the host Nicotiana benthamiana, and the defensin protein WT1 can be purified from virus-infected leaves by heat treatment and affinity chromatography. WT1 exhibits strong antifungal activity toward the phytopathogenic fungi Magnaporthe grisea(50% inhibitory concentration [IC50] = 5 μg/ml) and Botrytis cinerea (IC50 = 20 μg/ml) but is weakly active against the phytopathogenic bacterium Pseudomonas cichorii. This virus-mediated expression system is a rapid and efficient method to produce and characterize antimicrobial proteins in plants. It is particularly useful for the study of proteins that are difficult to produce with other expression systems.https://apsjournals.apsnet.org/doi/10.1094/MPMI.2001.14.2.111degenerate primerHis6immunoblot analysisPCRSDS-PAGE |
| spellingShingle | Hiromasa Saitoh Akinori Kiba Masahiro Nishihara Saburo Yamamura Kazumi Suzuki Ryohei Terauchi Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector Molecular Plant-Microbe Interactions degenerate primer His6 immunoblot analysis PCR SDS-PAGE |
| title | Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector |
| title_full | Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector |
| title_fullStr | Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector |
| title_full_unstemmed | Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector |
| title_short | Production of Antimicrobial Defensin in Nicotiana benthamiana with a Potato Virus X Vector |
| title_sort | production of antimicrobial defensin in nicotiana benthamiana with a potato virus x vector |
| topic | degenerate primer His6 immunoblot analysis PCR SDS-PAGE |
| url | https://apsjournals.apsnet.org/doi/10.1094/MPMI.2001.14.2.111 |
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