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A kind of extraction method of non-denatured protein and its special extracting solution

An extraction method and a non-denaturing technology, which are applied in the field of non-denaturing plant protein extraction and its special extraction solution, can solve the problems of inability to meet the non-denatured protein extraction requirements of Populus euphratica leaves, increasing the risk of dissolving pigment molecules, and interfering with electrophoresis patterns. , to achieve the effect of great application value, mild reagents, and efficient extraction

Active Publication Date: 2020-08-11
MINZU UNIVERSITY OF CHINA
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Problems solved by technology

Since the existing traditional protein extraction methods cannot meet the requirements for the extraction of non-denatured proteins from Populus euphratica leaves, especially the research on membrane proteins that require protein solubilization is even less applicable, and the introduction of non-ionic detergents will increase the pigment molecules at the same time. Risk of dissolution, severely interferes with electropherograms

Method used

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  • A kind of extraction method of non-denatured protein and its special extracting solution
  • A kind of extraction method of non-denatured protein and its special extracting solution
  • A kind of extraction method of non-denatured protein and its special extracting solution

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Embodiment

[0049] Example, comparison of 5 non-denatured protein extraction methods

[0050] The inventor of the present invention collected whole and insect-free fresh Populus euphratica leaves from the natural Populus euphratica forest (101°16'54.60"E, 42°2'35.67"N) from Badaoqiao, Ejina Banner, Alxa League, Inner Mongolia, and then quickly frozen in liquid nitrogen. A sample of Populus euphratica leaves was obtained.

[0051] 1. Use 5 methods to extract the non-denatured protein of Populus euphratica leaf samples

[0052] A, imidazole method

[0053] 1. Place 1g of Populus euphratica leaf sample in a mortar, add 100mg PVPP and a small amount of quartz sand, and grind thoroughly with liquid nitrogen to obtain powder.

[0054] 2. After completing step 1, place the powder in another new mortar and add 3 mL of extract to homogenize thoroughly to obtain a homogenized solution. The extract is a 0.001 mol / L EDTA buffer containing 0.05 mol / L imidazole, 0.002 mol / L 6-aminoacetic acid and 3% (3 mg / 100 ...

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Abstract

The invention discloses an extraction method of non-denatured protein and extraction liquid specially used by the method. The extraction method comprises a step a2): extracting the non-denatured protein in a sample by adopting the extraction liquid, wherein the extraction liquid contains sucrose, NaCl, L-sodium ascorbate, digitonin, dithiothreitol, phenylmethylsulfonyl fluoride and a protease inhibitor. The extraction method provided by the invention is easy and convenient to operate, a reagent is mild and economic, high-quality non-denatured protein can be efficiently extracted, the method lays a foundation and provides technical support for subsequently performed adverse plant proteomics research, and the method has a certain guiding significance for extracting recalcitrant tissue protein of plants living in an extreme environment. The method has a significant application value.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method for extracting non-denatured protein and a special extract solution thereof, in particular to a method for extracting non-denatured plant protein and a special extract solution thereof. Background technique [0002] As the main bearer of life activities, proteins mostly exist in the form of protein complexes and interacting proteins in cells, and their structures and functions are very complex. Traditional two-dimensional electrophoresis can only distinguish the denatured proteins extracted from plants, not their state when they assume functions in cells, and is not conducive to the study of protein-protein interactions and protein complexes. Protein separation is the core of electrophoresis technology, and a good protein extraction method is the prerequisite and guarantee for obtaining intuitive protein visualization information. Up to now, the traditional methods fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14
CPCC07K1/145
Inventor 王晓东冯金朝石莎韩航
Owner MINZU UNIVERSITY OF CHINA
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