Keywords

Multiple Enzyme Cutting Sites, Yellow Fluorescent Protein, Universal Vector, Laser Confocal, Plant Gene Characterization Tools

Abstract

The plant expression vector pCAMBIA3301 is used as the basic skeleton, and yellow fluorescent protein (YFP) is used as the tag protein to construct an expression vector that can fuse the target protein and contains a nuclease enzyme with a single recognition site for exogenous gene insertion. Cutting sites (Spe ¢ñ, Xba ¢ñ, Sma ¢ñ, BamH ¢ñ). In order to verify the practicability of the vector, the constructed vector was transformed into competent GV3101 Agrobacterium, colony PCR was identified, and then the tobacco lower epidermis was transiently transformed and Arabidopsis thaliana was stably transformed. The results of laser confocal microscopy showed that fluorescence was observed in the positive transgenic plants, but no fluorescence was observed in the negative control, indicating that the YFP-tagged protein can be expressed normally in the transgenic recipient cells. The successful construction of pCAMBIA3301::YFP vector provides a stable and reliable universal vector resource for research on plant protein subcellular localization and overexpression of transgenic plants and other related fields.

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Original research done by Kuang Chen, Zhu Xiaoyi, Zhang Liang, Fan Shihang, Hua Wei

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