Gateway-compatible vectors for functional analysis of proteins in cell type specific manner
Abstract Background Genetically encoded fluorescent proteins are often used to label proteins and study protein function and localization in vivo. Traditional cloning methods mediated by restriction digestion and ligation are time-consuming and sometimes difficult due to the lack of suitable restric...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-07-01
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| Series: | Plant Methods |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13007-020-00635-z |
| _version_ | 1818932610411266048 |
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| author | Liu Zhang Yang Zhao Haiyan Liang Xugang Li Kimberly L. Gallagher Shuang Wu |
| author_facet | Liu Zhang Yang Zhao Haiyan Liang Xugang Li Kimberly L. Gallagher Shuang Wu |
| author_sort | Liu Zhang |
| collection | DOAJ |
| description | Abstract Background Genetically encoded fluorescent proteins are often used to label proteins and study protein function and localization in vivo. Traditional cloning methods mediated by restriction digestion and ligation are time-consuming and sometimes difficult due to the lack of suitable restriction sites. Invitrogen developed the Gateway cloning system based on the site-specific DNA recombination, which allows for digestion-free cloning. Most gateway destination vectors available for use in plants employ either the 35S or ubiquitin promoters, which confer high-level, ubiquitous expression. There are far fewer options for moderate, cell-type specific expression. Results Here we report on the construction of a Gateway-compatible cloning system (SWU vectors) to rapidly tag various proteins and express them in a cell-type specific manner in plants. We tested the SWU vectors using the HISTONE (H2B) coding sequence in stable transgenic plants. Conclusions The SWU vectors are a valuable tool for low cost, high efficiency functional analysis of proteins of interest in specific cell types in the Arabidopsis root. |
| first_indexed | 2024-12-20T04:35:13Z |
| format | Article |
| id | doaj.art-01660badc7b743babe67d01c05876e42 |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-20T04:35:13Z |
| publishDate | 2020-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-01660badc7b743babe67d01c05876e422022-12-21T19:53:16ZengBMCPlant Methods1746-48112020-07-0116111010.1186/s13007-020-00635-zGateway-compatible vectors for functional analysis of proteins in cell type specific mannerLiu Zhang0Yang Zhao1Haiyan Liang2Xugang Li3Kimberly L. Gallagher4Shuang Wu5College of Life Sciences, FAFU-UCR Joint Center and Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, Fujian Agriculture and Forestry UniversityCollege of Life Sciences, FAFU-UCR Joint Center and Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, Fujian Agriculture and Forestry UniversityCollege of Resource and Environment, Fujian Agriculture and Forestry UniversityState Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural UniversityUniversity of PennsylvaniaCollege of Life Sciences, FAFU-UCR Joint Center and Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, Fujian Agriculture and Forestry UniversityAbstract Background Genetically encoded fluorescent proteins are often used to label proteins and study protein function and localization in vivo. Traditional cloning methods mediated by restriction digestion and ligation are time-consuming and sometimes difficult due to the lack of suitable restriction sites. Invitrogen developed the Gateway cloning system based on the site-specific DNA recombination, which allows for digestion-free cloning. Most gateway destination vectors available for use in plants employ either the 35S or ubiquitin promoters, which confer high-level, ubiquitous expression. There are far fewer options for moderate, cell-type specific expression. Results Here we report on the construction of a Gateway-compatible cloning system (SWU vectors) to rapidly tag various proteins and express them in a cell-type specific manner in plants. We tested the SWU vectors using the HISTONE (H2B) coding sequence in stable transgenic plants. Conclusions The SWU vectors are a valuable tool for low cost, high efficiency functional analysis of proteins of interest in specific cell types in the Arabidopsis root.http://link.springer.com/article/10.1186/s13007-020-00635-zFluorescent proteinsPhotoconversionGateway vectorsCell-specific expressionArabidopsis |
| spellingShingle | Liu Zhang Yang Zhao Haiyan Liang Xugang Li Kimberly L. Gallagher Shuang Wu Gateway-compatible vectors for functional analysis of proteins in cell type specific manner Plant Methods Fluorescent proteins Photoconversion Gateway vectors Cell-specific expression Arabidopsis |
| title | Gateway-compatible vectors for functional analysis of proteins in cell type specific manner |
| title_full | Gateway-compatible vectors for functional analysis of proteins in cell type specific manner |
| title_fullStr | Gateway-compatible vectors for functional analysis of proteins in cell type specific manner |
| title_full_unstemmed | Gateway-compatible vectors for functional analysis of proteins in cell type specific manner |
| title_short | Gateway-compatible vectors for functional analysis of proteins in cell type specific manner |
| title_sort | gateway compatible vectors for functional analysis of proteins in cell type specific manner |
| topic | Fluorescent proteins Photoconversion Gateway vectors Cell-specific expression Arabidopsis |
| url | http://link.springer.com/article/10.1186/s13007-020-00635-z |
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