Method for detecting lateral limitation area of living plant cytomembrane protein

A technology of laterally restricted area and cell membrane, which is applied in the field of detection of laterally restricted area of ​​living plant cell membrane proteins, can solve the problems that have not been clearly reported, and achieve the effect of simple method and good repeatability

Inactive Publication Date: 2015-09-09
INST OF BOTANY CHINESE ACAD OF SCI
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  • Abstract
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Problems solved by technology

However, how to achieve this purpose has not been clearly reported in the prior art

Method used

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  • Method for detecting lateral limitation area of living plant cytomembrane protein
  • Method for detecting lateral limitation area of living plant cytomembrane protein
  • Method for detecting lateral limitation area of living plant cytomembrane protein

Examples

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Embodiment 1

[0038] Example 1. Detection of the laterally restricted area of ​​phototropin phot1 on the cell membrane in Arabidopsis

[0039] The protein used to detect the laterally restricted area on the membrane is the Arabidopsis phototropin phot1, which is expressed on the epidermal cell membrane of the Arabidopsis hypocotyl. The phot1 tag was tagged using the green fluorescent protein GFP.

[0040] 1. Plant material

[0041] 1. Obtaining transgenic Arabidopsis

[0042] The Arabidopsis thaliana transformed into the fusion gene phot1-GFP under the phot1-5 mutant background used in the present invention comes from the document "Sakamoto and Briggs. Cellular and subcellular localization of phototropin 1. Plant Cell, 2002, 14:1723-1735" Publicly available and available to the public from the Institute of Botany, Chinese Academy of Sciences.

[0043] 2. Cultivation of Arabidopsis seeds

[0044] Transgenic Arabidopsis seeds were placed in a 1.5ml centrifuge tube, soaked in 2.5% (volume ...

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Abstract

The invention discloses a method for detecting the lateral limitation area of a living plant cytomembrane protein. The method comprises the following steps of (1) enabling a plant to express a fusion protein, so as to obtain a transgenic plant material which performs monochrome fluorescently labeling on the to-be-detected protein; (2) observing the transgenic plant material by using a variable-angle total internal reflection fluorescence microscope, so as to obtain a time sequence image; (3) recognizing and tracing a phosphor dot in a time series sequential image by using a single particle tracer technique, so as to obtain the trace of the phosphor dot along with the time and further calculate the mean square displacement of the phosphor dot along with the time; (4) fitting the mean square displacement of the phosphor dot along with the time by using Origin software, so as to obtain the corresponding constant, and calculating the moving range of the phosphor dot. The method for calculating the lateral limitation area of the living plant cytomembrane protein is simple, has good repeatability, can calculate the individual information of the protein and can calculate the population mean information.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for detecting the lateral confinement area of ​​living plant cell membrane proteins. Background technique [0002] Many important functions of the plasma membrane are closely related to the fluidity of the membrane. As an important part of the plasma membrane, membrane proteins can undergo lateral diffusion and endocytosis cycles, and play a key role in signal transmission and material transport through these movements. Therefore, detecting the distribution and diffusion movement of membrane proteins has become an important content in the study of biofilms. However, this rapid microsecond-scale motion on the nanoscale membrane structure cannot be observed with conventional microscope imaging. [0003] Through fluorescent labeling, the use of high-resolution single-molecule imaging technology can realize in vivo imaging of the dynamics of membrane protein molecules at the nan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12N15/82
Inventor 林金星薛轶群王晓华
Owner INST OF BOTANY CHINESE ACAD OF SCI
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