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Plant plasma membrane fluorescence labeling method and plasma membrane protein positioning and dynamic monitoring method

A fluorescent labeling and plasma membrane technology, applied in the field of plasma membrane staining, can solve the problems of complicated operation, long cycle and high difficulty, and achieve the effect of saving experimental time

Pending Publication Date: 2022-03-25
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Description
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  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows researchers to study plant cells by adding specific molecules called FM 46-464 (a special type of cyanine) dyes that are able to attach themselves on certain parts of their cell walls when exposed to light. These colored particles move along the entirety of the outer layer of the cell wall while carrying water into another part where they don't get stuck well enough. By doing this experimentally, scientists may investigate how different types of chemicals affect these structures inside them more easily than before studying other aspects like DNA replication.

Problems solved by technology

This patented technical problem addressed in this patent relates to developing a simpler and faster way for studying plasmas (a type of fluid containing charged particles) membranes without having complex operations or requiring expensive equipment such as lasers.

Method used

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  • Plant plasma membrane fluorescence labeling method and plasma membrane protein positioning and dynamic monitoring method
  • Plant plasma membrane fluorescence labeling method and plasma membrane protein positioning and dynamic monitoring method

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Embodiment 1

[0034] Material: transgenic Arabidopsis thaliana with plasma membrane localization protein DsSWEET12 (GenBank accession number: MG737823) (Zhou et al, A Novel Sugar Transporter from Dianthus spiculifolius, DsSWEET12, Affercts Sugar Metabolism and Confers Osmotic and Oxidative Stress Tolerance in Arabidlar Sicis. Molecu. 2018)

[0035] 1. Arabidopsis root hair cell plasma membrane staining;

[0036] Step 1. The transgenic Arabidopsis seeds were sterilized with 70% alcohol for 30 seconds, sterilized with sodium hypochlorite for 3 minutes, washed with sterile water for 5 times, planted on solid 1 / 2 MS medium, and placed at 4°C for 2 days;

[0037] Step 2. Take out the petri dish with seeds and place it vertically in a light incubator for 7 days. The culture temperature is 22°C, the photoperiod is 12h light / 12h dark, and the light intensity is 150μE m -2 the s -1 ;

[0038] Step 3: Use scissors to cut the whole root of Arabidopsis seedlings grown vertically for 7 days in a petr...

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Abstract

The invention provides a plasma membrane fluorescence labeling method of a plant and a plasma membrane protein positioning and dynamic monitoring method, and belongs to the technical field of plasma membrane dyeing. According to the invention, the FM4-64 dye is adopted to dye the plasma membrane structure, and the FM4-64 dye can be embedded in the plasma membrane bimolecular layer and can be dynamically transferred along with the plasma membrane, so that the FM4-64 dye can also be used by subsequent related researches such as endocytosis and transfer of the plasma membrane. And moreover, the fluorescence labeling method disclosed by the invention can be used for carrying out fluorescence labeling on the plasma membrane during genetic transformation or instantaneous transformation, so that the experiment time is saved, and the interference problem of a fluorescence signal does not exist.

Description

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Claims

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Application Information

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Owner NORTHEAST AGRICULTURAL UNIVERSITY
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