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Bidirectional electrophoresis method for malus plant protein

A plant protein and two-dimensional electrophoresis technology, which is applied in peptide preparation methods, chemical instruments and methods, organic chemistry, etc., can solve the problems of lagging proteome research and large interference of two-dimensional electrophoresis, and achieve the effect of reducing interference

Inactive Publication Date: 2008-09-10
CHINA AGRI UNIV
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Problems solved by technology

This is due to the polysaccharides, phenols and nucleic acid substances in the plants of the small golden crabapple, the existence of these substances greatly interferes with the two-dimensional electrophoresis, resulting in a relatively lagging behind in the proteome research related to the plants of the genus Malus

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  • Bidirectional electrophoresis method for malus plant protein

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Embodiment 1

[0017] Embodiment 1, protein two-dimensional electrophoresis of Malus xiaojinensis

[0018] The protein two-dimensional electrophoresis of the present invention includes the steps of sample preparation, protein quantification, two-dimensional gel electrophoresis, protein staining and gel image analysis, and the specific methods are as follows:

[0019] 1. Sample Preparation

[0020] 1) Put 2 g of the young white primary root system of Malus xiaojinensis Cheng et. Jiang (the number in the National Apple Germplasm Resource Garden is DGB0458) in an ice bath in liquid nitrogen and grind until it becomes a white powder;

[0021] 2) Add 2ml-20°C pre-cooled acetone solution containing 10% trichloroacetic acid and 0.07% β-mercaptoethanol to continue grinding to a homogenate, transfer to a centrifuge tube, place at -20°C for 1h, then 4°C Centrifuge at 12000rpm for 15min, discard the supernatant;

[0022] 3) Add 4 times the volume of the precipitate to the acetone solution containing ...

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Abstract

The invention discloses a dielectrophoresis method for proteins of malus plants. The steps of the method are as follows: 1) plant materials are ground in liquid nitrogen and are added with an acetone solution which is pre-cooled at a temperature of 20 DEG C below zero and contains a trichloroacetic acid with the concentration of 10g / 100ml and beta-mercaptoethanol with the concentration of 0.07ml / 100ml and continuously ground to be the homogenate, and the homogenate stands at a temperature of 20 DEG C below zero for 1 hour and centrifuged by the rotary speed of 12000rpm for 15 minutes, and the precipitation is kept; 2) the precipitation is added with an acetone solution which contains the beta-mercaptoethanol with the concentration of 0.07ml / 100ml and the volume of which is 4 times of the volume of the precipitation, and the mixed solution is evenly mixed and stands at a temperature of 20 DEG C below zero for 1 hour, and centrifuged by the rotary speed of 12000rpm for 15 minutes, and the precipitation is kept; 3) the step 2 is repeated for 6 times, and total proteins are obtained; 4) total proteins obtained from the step 3 undergo the electric focusing electrophoresis, and the sample volume of proteins is of 300ug / 170mm adhesive tape, and the polyacrylamide gel electrophoresis is conducted, and the silver dyeing is conducted after the electrophoresis.

Description

technical field [0001] The invention relates to a two-dimensional electrophoresis method for malic plant protein. Background technique [0002] Iron is one of the essential trace elements for plant growth and development. Using biological technology to solve the problem of iron deficiency in plants is an effective method. Little golden crabapple is the world's first iron-deficiency-tolerant apple plant. It is particularly important to study the stress-resistant mechanism of small golden crabapple for cultivating stress-resistant fruit tree varieties. Proteome analysis is an important technical means to study the stress response of plants, and it is helpful to study the mechanism of iron deficiency tolerance in plants. Protein two-dimensional electrophoresis is a commonly used technical method in proteome analysis. Up to now, there is no two-dimensional electrophoresis analysis method suitable for Malus plants. This is due to the polysaccharides, phenols and nucleic acid su...

Claims

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Application Information

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IPC IPC(8): C07K1/26
Inventor 韩振海王晶莹许雪峰李天忠王忆
Owner CHINA AGRI UNIV
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