Method for efficiently extracting sheep viscus tissue protein

A tissue protein, sheep technology, applied in the field of sheep visceral tissue protein extraction, can solve the problems of high technical requirements and complicated operations, achieve high purity, less insoluble protein, and demonstrate the effect of technological progress

Inactive Publication Date: 2014-05-07
新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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Problems solved by technology

② The patent No. CN 101252849A's "System and Method for Separating Proteins from Connective Tissue" discloses the separation of muscle proteins from connective tissue, and the generation of slurry by placing animal tissues including muscle tissue and connective tissue in an aqueous solvent. The slurry is then accelerated in a low-shear environment, enabling the separation of muscle proteins from connective tissue, but the method is complex and technically demanding

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  • Method for efficiently extracting sheep viscus tissue protein
  • Method for efficiently extracting sheep viscus tissue protein
  • Method for efficiently extracting sheep viscus tissue protein

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Embodiment

[0025] The rapid and economical method for extracting sheep visceral tissue protein provided by the present invention uses sheep liver, spleen, lung, kidney and other visceral tissues as raw materials, including sample pretreatment, cell disruption, protein quantification, and protein detection (SDS-PAGE and Western-Blot) consists of several steps, the specific method is as follows:

[0026] 1) Weigh 25 mg of different visceral tissue samples and place them in a 2 mL centrifuge tube that has been cooled in advance, and use autoclaved scissors to cut the samples into small pieces of 3 mm × 3 mm;

[0027] 2) Add pre-cooled lysate containing cocktail protease inhibitor at 4% volume before use. The ratio of sample to lysate is 1:3. At the same time, add sterile steel balls with a diameter of 5 cm and place it in the TissueLyser LT instrument of QIAGEN. Homogenate (pre-cool the instrument in a -70 refrigerator before use), for visceral tissue homogenate for 50 seconds, until the sa...

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Abstract

The invention provides a method for efficiently extracting sheep viscus tissue protein, i.e. a frozen crushing homogenate method and a vortex mixing method are adopted, the protein quantification is carried out on the extracted viscus tissue protein of livers, spleens, lungs, kidneys and the like of the sheep, the extracted protein is separated through polyacrylamide gel electrophoresis, and the expression of the transgenosis sheep green fluorescent protein (GFP) genes is detected by western blot. The method has the technical characteristics that: 1, the concentration of the extracted protein is high, and the cost is low; 2, the method is applicable to the extraction of viscus tissue protein of adult or young sheep; and 3, insoluble protein is little, the abandoned tissue residue is little, and important practical values are realized.

Description

technical field [0001] The invention relates to the extraction of sheep visceral tissue protein, which combines the freezing crushing homogenization method and the vortex mixing method to efficiently extract the protein in the sheep visceral tissue, and verifies its important practical value by expressing the green fluorescent protein gene. Background technique [0002] Animal visceral tissue is the site for the synthesis and metabolism of various biologically active substances, and contains a variety of proteins. It is often used to detect the expression and expression of biologically active substances and certain proteins. Animal proteins generally exist in the form of complex mixtures in tissues or cells, and each type of cell contains thousands of different proteins, and its extraction and purification is a difficult and complicated task; from the complex mixture There is no universally applicable method to completely extract a certain animal protein; but any animal pro...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14G01N33/68
Inventor 张雪梅李文蓉贺三刚张宁刘明军
Owner 新疆维吾尔自治区畜牧科学院中国-澳大利亚绵羊育种研究中心
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