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

• Main Authors: Liu Zhang, Yang Zhao, Haiyan Liang, Xugang Li, Kimberly L. Gallagher, Shuang Wu
• Format: Article
• Language: English
• Published: BMC 2020-07-01
• Series: Plant Methods
• Subjects: Fluorescent proteins; Photoconversion; Gateway vectors; Cell-specific expression; Arabidopsis
• Online Access: http://link.springer.com/article/10.1186/s13007-020-00635-z

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.