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Apple protein extraction method and related protein extracting solution

An extraction method, protein technology, applied in the direction of peptide preparation methods, chemical instruments and methods, material inspection products, etc., can solve complex protein extraction problems

Inactive Publication Date: 2016-08-17
FRUIT TREE INST OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction of protein is the key and basis of the quantitative analysis of iTRAQ markers; compared with other model plants, apple leaf tissue contains a large amount of secondary metabolites such as phenols and tannins, making the extraction of protein more complicated

Method used

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  • Apple protein extraction method and related protein extracting solution
  • Apple protein extraction method and related protein extracting solution
  • Apple protein extraction method and related protein extracting solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, extraction and identification of apple protein

[0048] 1. Apple protein extraction

[0049] Extract apple protein as follows:

[0050]1) Pick apple leaves with a leaf age of 20-25 days ("Huahong"), wash them with sterile water, and then quickly air-dry them; add 0.2 grams of polyvinylpolypyrrolidone to 2 grams of apple leaves to obtain a mixture, which is named as mixture 1 ; The mixture 1 is quickly ground in a mortar with liquid nitrogen to obtain plant powder;

[0051] 2) Add 5ml of protein extract to the plant powder obtained in step 1) and vortex at 4°C for 5 minutes to obtain an extract containing protein, which is called extract 1; An equal volume of Tris-saturated phenol in extract 1 was vortexed and shaken for 5 minutes to obtain an extract containing protein, which was named extract 2; Enter the phenolic phase, collect the phenolic phase, add 100mM methanolic ammonium acetate solution 5 times the volume of the phenolic phase to the phenolic p...

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Abstract

The invention discloses an apple protein extraction method and a related protein extracting solution. The disclosed protein extraction method includes the steps that plant tissue and / or organs are smashed, the protein extracting solution is added and reacts for 3-7 minutes at the temperature of 0-6 DEG C, impurities are removed to obtain protein; solvent of the protein extracting solution is water, the protein extracting solution contains solutes of 100 mM of tri-hydroxymethyl aminomethane, 50 mM of ethylenediaminetetraacetic acid, 50 mM of potassium chloride, 25 mM of sodium tetraborate, 1 mM of dithiothreitol, 2 mM of phenylmethylsulfonyl fluoride, 1%(volume percentage concentration) polyethylene glycol octylphenol ether, 5% (volume percentage concentration) beta-mercaptoethanol and saccharose of 25 mg / mL, and the pH value is 7.5-8.0. By means of the protein extraction method, impurities such as polyphenol and pigment can be effectively removed, and when obtained protein is quantitatively analyzed through an iTRAQ technology, the number of the obtained protein can be larger than 2000.

Description

technical field [0001] The invention relates to a method for extracting apple protein and a related protein extract in the field of biotechnology. Background technique [0002] At present, with the continuous update and improvement of molecular biology technology, molecular biology methods also provide convenient conditions for apple genetic research; as early as 1989, the European Apple Genome Project was launched, and apple genetic research was its long-term focus In recent years, proteomics research with the advantages of high throughput and high sensitivity has become the frontier and hot field of life science research. [0003] At present, with the continuous deepening and improvement of fruit tree genome research, proteomic analysis technology has been applied to the research of apple, pear, grape, peach, banana and other fruit tree varieties. In 2010, the draft genome of the apple cultivar "Golden Delicious" was published. The massive data generated by the sequencing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/30C07K1/14G01N33/68
CPCC07K1/30C07K1/14G01N33/6848
Inventor 张彩霞田义张利义丛佩华
Owner FRUIT TREE INST OF CHINESE ACAD OF AGRI SCI
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