4.6 Article

Gateway-compatible vectors for functional analysis of proteins in cell type specific manner

Journal

PLANT METHODS
Volume 16, Issue 1, Pages -

Publisher

BMC
DOI: 10.1186/s13007-020-00635-z

Keywords

Fluorescent proteins; Photoconversion; Gateway vectors; Cell-specific expression; Arabidopsis

Funding

  1. National Natural Science Foundation of China [31722006]
  2. National Key Research and Development Program of China [2016YFD0100700]

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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 theArabidopsisroot.

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