Extraction method of non-denatured protein and extraction liquid specially used by method

An extraction method and technology of extracting solution, which are applied in the field of non-denatured plant protein extraction method and its special extracting solution, can solve the problems of not being able to meet the requirements for the extraction of non-denatured protein from Populus euphratica leaves, increasing the risk of pigment molecule dissolution, and interfering with electrophoretic spectra. , to achieve significant application value, mild reagents, and efficient extraction

Active Publication Date: 2018-05-18
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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  • Extraction method of non-denatured protein and extraction liquid specially used by method
  • Extraction method of non-denatured protein and extraction liquid specially used by method
  • Extraction method of non-denatured protein and extraction liquid specially used by method

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Embodiment

[0049] The comparison of the extraction method of embodiment, 5 kinds of non-denatured proteins

[0050] The inventor of the present invention collects complete fresh Populus euphratica leaves without insect bites from the natural Populus euphratica forest (101 ° 16'54.60 "E, 42 ° 2' 35.67 "N) in Badaoqiao, Ejina Banner, Alxa League, Inner Mongolia, and then quickly freezes them in liquid nitrogen. Get samples of Populus euphratica leaves.

[0051] 1. Five methods were used to extract non-denatured proteins from Populus euphratica leaf samples

[0052] A, imidazole method

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

[0054] 2. After step 1 is completed, put the powder in another new mortar, add 3mL extract solution to fully homogenize, and obtain a homogenate solution. The extract is 0.001mol / L EDTA buffer solution containing 0.05mol / L imidazole,...

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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 in particular relates to a method for extracting non-denatured protein and a special extract thereof, in particular to a method for extracting non-denatured plant protein and a special extract 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 denatured proteins extracted from plants, not their state when they are functional in cells, which 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 premise and guarantee for obtaining intuitive protein visualization information. Up to now, the traditional method of extracting non-denatu...

Claims

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Application Information

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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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