Method for extracting cyst-like protein

An extraction method, protein technology, applied in the preparation method of peptides, chemical instruments and methods, peptides, etc.

Pending Publication Date: 2020-05-26
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no problem with the method of extracting thylakoid protei...

Method used

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  • Method for extracting cyst-like protein
  • Method for extracting cyst-like protein

Examples

Experimental program
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Embodiment

[0039] 1) Experiment preparation: prepare an appropriate amount of grinding buffer GB (50mM HEPES / KOH, pH 7.5; 0.33Msorbitol; 1mM MgCl 2 ; 1mM MnCl 2 ; 2mM EDTA, pH8.0; 5mM Na-ascorbate; 0.1%BSA), resuspension buffer RB (50mM HEPES-KOH, pH 7.9; 0.33M sorbitol; 0.8mM CaCl 2 ; 5mM Na-ascorbate), 5×loading buffer (1M Tris / HCl,pH6.8; 50% glycerol; 10% SDS; 5%β-mercaptoethanol; 0.5% bromophenol blue) and 1×TE buffer (10mMTricine- KOH, pH7.5; 2mM EDTA); turn on the centrifuge, adjust the temperature to 4°C for pre-cooling; wash all kinds of centrifuge tubes, funnels and nylon mesh and blow dry.

[0040] 2) Use a juice extractor to extract chloroplasts: take 5g of leaves, cut them into about 80mL GB, then use a juice extractor to crush them, jog 3 times, 5s each time. Filter the resulting solution into a large round-bottom centrifuge tube, centrifuge at 2000×g for 1-2 min, discard the supernatant, and add 500 μL RB to resuspend the pellet. Carefully add the resuspension to the pre...

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Abstract

The invention belongs to the technical field of biological separation engineering, and particularly relates to a method for extracting cyst-like protein. At present, a method for extracting the cyst-like protein does not exist. The invention provides the method for extracting the cyst-like protein, which comprises the following steps: 1) cutting leaves into pieces, putting the pieces into a grinding buffer solution GB, and crushing; 2) separating chloroplast in the broken leaf cells, and obtaining the chloroplast with high purity and complete form; 3) labeling a sample to be subjected to component separation, and then centrifuging; 4) resuspending a sample for component separation by using a sucrose-containing 1*TE buffer, and cracking chloroplast; and 5) separating a supernatant from theprecipitate through centrifugation to obtain the cyst-like protein, which is mainly used for analyzing and researching important functions and action mechanisms of the cyst-like protein in various life activities, and has important biological significance for explaining physiological functions and genetic rules of the cyst-like protein.

Description

technical field [0001] The application belongs to the technical field of bioseparation engineering, in particular to a method for extracting thylakoid proteins. Background technique [0002] Thylakoids are light reaction sites for photosynthesis in plants, located in chloroplasts, and composed of grana and stroma. The grana lamellae are stacked up and down by many flat cystic structures with a diameter of about 400nm, and the granum lamellae are connected with the grana thylakoids. Chlorophyll and other pigments are distributed on the membranes of thylakoids. Various enzymes, proteins, photocomplexes PSⅠ, photocomplexes PSⅡ, etc. required for photosynthesis are distributed on the membrane or matrix of the grana. Thylakoid is an interconnected network structure, in which the complex folded membrane system greatly increases the photosynthetic area, thus improving the photosynthetic efficiency. [0003] Photosynthesis can convert active solar energy into stable chemical ener...

Claims

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Application Information

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IPC IPC(8): C07K1/14
CPCC07K1/145
Inventor 刘小敏王璐璐高宏波
Owner BEIJING FORESTRY UNIVERSITY
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