A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein
The bimolecular fluorescence complementation (BiFC) assay was developed as a tool for the visualization of protein–protein interactions in living cells. To date, many types of BiFC systems with distinct colors have been developed. Most of the colors in the visible spectrum have been used in BiFC ass...
| 出版年: | BioTechniques |
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| 主要な著者: | , , |
| フォーマット: | 論文 |
| 言語: | 英語 |
| 出版事項: |
Taylor & Francis Group
2018-04-01
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| 主題: | |
| オンライン･アクセス: | https://www.future-science.com/doi/10.2144/btn-2017-0121 |
| _version_ | 1849877129521528832 |
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| author | Yuta Fujii Ayaka Yoshimura Yutaka Kodama |
| author_facet | Yuta Fujii Ayaka Yoshimura Yutaka Kodama |
| author_sort | Yuta Fujii |
| collection | DOAJ |
| container_title | BioTechniques |
| description | The bimolecular fluorescence complementation (BiFC) assay was developed as a tool for the visualization of protein–protein interactions in living cells. To date, many types of BiFC systems with distinct colors have been developed. Most of the colors in the visible spectrum have been used in BiFC assays, with the exception of orange. In this study, we developed an orange-colored BiFC system using the Kusabira-Orange (KO) protein from the stony coral Fungia concinna. To obtain bright BiFC fluorescence, we compared fluorescence intensities of two monomeric KO variants (mKO1 and mKO2) and identified mKO2 as brighter than mKO1. The optimal split site for mKO2-based BiFC was defined by a comparative analysis of complementation efficiency and a signal-to-noise ratio. The resulting mKO2-based BiFC system successfully demonstrated protein dimerization in plant cells as a model experiment. The novel mKO2-based BiFC system will expand the possibility of multicolor BiFC analysis. |
| format | Article |
| id | doaj-art-e17c406526b24d30bcae89e5fc10974e |
| institution | Directory of Open Access Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2018-04-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| spelling | doaj-art-e17c406526b24d30bcae89e5fc10974e2025-08-20T01:11:21ZengTaylor & Francis GroupBioTechniques0736-62051940-98182018-04-0164415316110.2144/btn-2017-0121A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange proteinYuta Fujii0Ayaka Yoshimura1Yutaka Kodama2Center for Bioscience Research and Education, Utsunomiya University, Tochigi, JapanCenter for Bioscience Research and Education, Utsunomiya University, Tochigi, JapanCenter for Bioscience Research and Education, Utsunomiya University, Tochigi, JapanThe bimolecular fluorescence complementation (BiFC) assay was developed as a tool for the visualization of protein–protein interactions in living cells. To date, many types of BiFC systems with distinct colors have been developed. Most of the colors in the visible spectrum have been used in BiFC assays, with the exception of orange. In this study, we developed an orange-colored BiFC system using the Kusabira-Orange (KO) protein from the stony coral Fungia concinna. To obtain bright BiFC fluorescence, we compared fluorescence intensities of two monomeric KO variants (mKO1 and mKO2) and identified mKO2 as brighter than mKO1. The optimal split site for mKO2-based BiFC was defined by a comparative analysis of complementation efficiency and a signal-to-noise ratio. The resulting mKO2-based BiFC system successfully demonstrated protein dimerization in plant cells as a model experiment. The novel mKO2-based BiFC system will expand the possibility of multicolor BiFC analysis.https://www.future-science.com/doi/10.2144/btn-2017-0121BiFC competition assayGateway cloningKO proteinmulticolor BiFCpGWT35Sprotein interaction |
| spellingShingle | Yuta Fujii Ayaka Yoshimura Yutaka Kodama A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein BiFC competition assay Gateway cloning KO protein multicolor BiFC pGWT35S protein interaction |
| title | A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein |
| title_full | A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein |
| title_fullStr | A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein |
| title_full_unstemmed | A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein |
| title_short | A novel orange-colored bimolecular fluorescence complementation (BiFC) assay using monomeric Kusabira-Orange protein |
| title_sort | novel orange colored bimolecular fluorescence complementation bifc assay using monomeric kusabira orange protein |
| topic | BiFC competition assay Gateway cloning KO protein multicolor BiFC pGWT35S protein interaction |
| url | https://www.future-science.com/doi/10.2144/btn-2017-0121 |
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