Method for removing RuBisco protein in broussonetia papyrifera leaf protein

A protein and protein technology, applied in the field of protein separation and detection, can solve problems such as limiting the detection of low-abundance proteins

Inactive Publication Date: 2016-06-08
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In two-dimensional gel electrophoresis (2-DE), the ultra-high abundance protein RuBisco will severely limit the detection of low abundance proteins

Method used

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  • Method for removing RuBisco protein in broussonetia papyrifera leaf protein
  • Method for removing RuBisco protein in broussonetia papyrifera leaf protein
  • Method for removing RuBisco protein in broussonetia papyrifera leaf protein

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Experimental program
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Effect test

Embodiment 1

[0057] Embodiment 1, remove RuBisco protein and effect detection in the leaf protein of mulberry

[0058] 1. Removal of RuBisco protein from mulberry leaf protein

[0059] 1. Prepare homogenate

[0060] Put 0.5 g of the leaves of the tree mulberry into 600 μL of GMI homogenate and grind to obtain the first homogenate, add 600 μL of GMII homogenate into the first homogenate, and grind evenly to obtain the second homogenate.

[0061] The solvent of the GMI homogenate is a 500mM Tris-HCl buffer solution (prepared with deionized water) with a pH of 8, and the solute and concentration are: 20mM MgCl 2 and 1mMPMSF.

[0062] The solvent of the GMⅡ homogenate is a 500mM Tris-HCl buffer solution (prepared with deionized water) with a pH of 8, and the solute and concentration are: 20mM MgCl 2 , 1mMPMSF and 4% (volume percentage) NonidetP-40.

[0063] 2. Homogenate centrifugation

[0064] Under the condition of 10000g, the second homogenate was centrifuged at 4° C. for 10 min, and t...

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Abstract

The invention discloses a method for removing RuBisco protein in broussonetia papyrifera leaf protein. The method can be used for removing RuBisco protein in all leaf proteins and comprises the following steps: (1) preparing homogenate; (2) centrifuging the homogenate; (3) centrifuging first supernate; (4) centrifuging second supernate; and (5) washing coarse protein precipitate. The method can be used for extracting leaf proteins of plants, reducing interference of high-abundant protein RuBisco, improving 2-DE electrophoresis resolution ratio and laying the foundation for optimizing detection of the leaf proteins of the plants and further improving varieties by using biotechnological manners.

Description

technical field [0001] The invention belongs to the field of protein separation and detection, and relates to a method for removing RuBisco protein from mulberry leaf protein. Background technique [0002] Paper mulberry is a multifunctional and comprehensive tree species, and the whole plant has development potential. Its leaves are rich in nutrients such as crude protein, crude fat and crude fiber, which can partially replace conventional feed such as soybean meal and wheat bran, and can be used for livestock breeding such as pigs, cattle and sheep. The formula technology structure of my country's feed industry is still dominated by corn-soybean meal. Bulk raw materials such as soybean meal and corn are more and more dependent on foreign countries, and are increasingly affected by the international market. Therefore, how to break through the existing formula technology and develop new raw materials and substitutes has become an important challenge to enhance the comprehe...

Claims

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Application Information

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IPC IPC(8): C07K1/30
CPCC07K1/30
Inventor 沈世华王小曼邳植藤林宏彭献军
Owner INST OF BOTANY CHINESE ACAD OF SCI
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