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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2020.1
- 수록면
- 131 - 138 (8page)
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Transient protein expression is a useful technique for investigating the protein localization and functional analysis of stress responses in rice plants. Currently, available methods for in planta transient expression analysis include Agrobacteriummediated transformation, protoplast transformation using polyethylene glycol or electroporation, and biolistic bombardment expression which have several disadvantages and are not well suited for the rice. Therefore, development of a method for rapid and efficient analysis of protein expression, subcellular localization, pathogen effector screening, and protein–protein interaction in rice is required. We developed a protocol for in planta gene expression analysis in sliced rice sheath cells by modifying and optimizing the biolistic particle bombardment technique. By obtaining thin sections (~400 μm) of rice sheath cells, auto-fluorescence from chlorophyll was eliminated. This system was validated through the localization of marker genes specifically expressed in nuclei, plasma membranes, and tonoplast. In addition, high transformation efficiency of 30% was achieved. Therefore, this protocol provides a new and rapid method for transient gene expression assay in rice. Protein secretion was examined in rice sheath cells using predicted secretory proteins from rice blast fungus, indicating that this method is applicable to plant–microbe interaction studies. The transient expression protocol established here is well optimized for protein localization, secretion, and host–pathogen protein interaction studies in rice. A typical experiment can be completed in three days.
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참고문헌 (37)
참고문헌 신청
Agrawal GK / 2010 / Plant secretome : unlocking secrets of the secreted proteins / Proteomics 10:799~827
Barros J / 2015 / The cell biology of lignification in higher plants / Ann Bot 115:1053~1074
Carsono N / 2008 / Transient expression of green fluorescent protein in rice calluses : optimization of parameters for helios gene gun device / Plant Prod Sci 11:88~95
Casasoli M / 2008 / Identification by 2-D DIGE of apoplastic proteins regulated by oligogalacturonides in Arabidopsis thaliana / Proteomics 8:1042~1054
Chivasa S / 2005 / Pathogen elicitor-induced changes in the maize extracellular matrix proteome / Proteomics 5:4894~4904
Barros J / 2015 / The cell biology of lignification in higher plants / Ann Bot 115:1053~1074
Carsono N / 2008 / Transient expression of green fluorescent protein in rice calluses : optimization of parameters for helios gene gun device / Plant Prod Sci 11:88~95
Casasoli M / 2008 / Identification by 2-D DIGE of apoplastic proteins regulated by oligogalacturonides in Arabidopsis thaliana / Proteomics 8:1042~1054
Chivasa S / 2005 / Pathogen elicitor-induced changes in the maize extracellular matrix proteome / Proteomics 5:4894~4904
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