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Plant cell lysis vacuole specific marker and application

A plant cell, specific labeling technology, applied in the field of molecular biology, can solve the problems of small number of vacuolar labeled proteins, inaccurate labeling affecting scientific research results, etc., and achieve strong specificity.

Active Publication Date: 2020-07-31
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, the number of vacuolar marker proteins is still very small, and the laboratory's preliminary research results show that when γ-TIP is used to mark lysed vacuoles in tobacco, part of the cell membrane can also be marked, so there is a hidden danger that inaccurate labeling will affect scientific research results. The specificity of the lytic vacuole can solve this problem well

Method used

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  • Plant cell lysis vacuole specific marker and application
  • Plant cell lysis vacuole specific marker and application
  • Plant cell lysis vacuole specific marker and application

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Experimental program
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Embodiment 1

[0030] 1. Acquisition of plant cell lysis vacuole-specific markers

[0031] 1. Collection of grape peronospora samples

[0032] Grape downy mildew usually begins to occur from June to July, and the peak period is from August to September. This is the best time for sampling. Such as figure 1 As shown, grape downy mildew mainly harms grape leaves, producing yellow to brown polygonal lesions, and a white mold layer on the back of the leaf spots. The disease samples were stored in ziplock bags after collection.

[0033] 2. Extraction of Peronospora grape RNA

[0034] (1) Take the diseased tissue (about 100 mg) containing Peronospora viticola in a 1.5 mL centrifuge tube, freeze it in liquid nitrogen and grind it into powder.

[0035] (2) Add 600 μL 65°C preheated RNA extraction solution (2% CTAB, 2% PVP-40, 2M NaCl, 20mM EDTA, 0.1M Tris-HCl, 1% β-mercaptoethanol, 0.5g / L Spermidine, pH 8.0 ), vortexed to mix evenly, and bathed in 65°C water for 30 minutes, and inverted twice du...

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Abstract

The invention discloses a specific marker for plant cell lysis vacuoles, and the specific marker for the plant cell lysis vacuoles is an RxLR132aa1-158 gene. The invention also discloses a protein coded by the specific marker and an application of the specific marker in fluorescence detection of plant lysate vesicles. A plurality of cyclic structures are marked by fluorescence of fusion protein expressed after the RxLR132aa1-158 is fused with a fluorescence reporter gene, and the fluorescence can be superposed with the fluorescence of the lysate labeled protein fusion protein, so that the lysate can be specifically labeled. In addition, specific positioning is not influenced by the fluorescence reporter gene, so that the specificity is stronger. The plant cell lysis vacuole specific markerenriches a lysis vacuole marker library, can provide various candidate marker selections for different plant protein lysis vacuole positioning researches, and has important significance.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a plant cell lysis vacuole specific marker and its application. Background technique [0002] Cells are the basic structural and functional units of living organisms. In eukaryotic cells, biomembranes separate eukaryotic cells into different subcellular structures, including mitochondria, lytic vacuoles, peroxisomes, lysosomes, endoplasmic reticulum, nucleus, Golgi apparatus, vacuoles, vesicles, etc. , so that multiple chemical reactions can be carried out simultaneously in the cell without interfering with each other. Various proteins are distributed orderly according to their functions in each compartment of the cell to function. Studying the subcellular localization of proteins is crucial for exploring and analyzing protein functions, and it is also an essential link for systematically understanding plant growth and development, morphogenesis, and resistance to var...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/37C12N15/65C12N15/82G01N21/64
CPCC07K14/37C12N15/65C12N15/8212G01N21/6486Y02A50/30
Inventor 卢江傅佩宁蓝霞吴伟宋士任高宇
Owner SHANGHAI JIAO TONG UNIV