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A method for detecting milk protein and milk fat synthesis ability of cow's mammary gland by using cow's milk

A technology of milk protein and milk fat, which is applied in the field of detection of milk protein and milk fat synthesis ability of dairy cow mammary gland, which can solve the problem of not accurately and comprehensively reflecting the ability of mammary gland to synthesize milk protein and milk fat

Inactive Publication Date: 2019-06-21
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the method for detecting milk protein and milk fat of raw milk at this stage cannot accurately and comprehensively reflect the ability of different dairy cow breeds and individual mammary glands to synthesize milk protein and milk fat, and provides a method for detecting milk protein and milk fat using milk. Method for Synthesizing Milk Protein and Milk Fat Ability of Dairy Cow Mammary Gland

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  • A method for detecting milk protein and milk fat synthesis ability of cow's mammary gland by using cow's milk
  • A method for detecting milk protein and milk fat synthesis ability of cow's mammary gland by using cow's milk
  • A method for detecting milk protein and milk fat synthesis ability of cow's mammary gland by using cow's milk

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specific Embodiment approach 1

[0013] Specific Embodiment 1: In this embodiment, a method for detecting milk protein and milk fat synthesis ability of cow mammary glands using cow milk: 1. Pretreatment of cow milk: take cow milk and freeze it at -80°C for later use to obtain milk samples; 2. Preparation of secretions: centrifuge milk samples at 0-4°C, 2000-2500rpm for 25-30min, remove precipitated cells and floating fat globules, and obtain skim milk; centrifuge skim milk at 0-4°C, 12000- Centrifuge at 13000rpm for 30-40min and take the supernatant; centrifuge the supernatant at 0-4°C and 100000-110000rpm for 2-2.5h to take the precipitate; dissolve the precipitate with 0.01-0.02M PBS to obtain the outer Primary exosome solution: centrifuge the primary exosome solution at 0-4°C, 100000-110000rpm for 2-2.5h, take the precipitate, dissolve the precipitate with 0.01-0.02M PBS solution, and obtain the exosome solution, Detect the purity of the exosome solution, and proceed to the next step after passing the tes...

specific Embodiment approach 2

[0026] Embodiment 2: This embodiment differs from Embodiment 1 in that: Western blotting is used to detect the purity of the exosome solution. Others are the same as the first embodiment.

[0027] In this embodiment, the steps for detecting the purity of the exosome solution by Western blotting are:

[0028] 1. Sample preparation: transfer the exosome solution to a 1.5ml centrifuge tube, add 2× loading buffer or 5× loading buffer, and then place in a boiling water bath for 10 minutes; ultrasonicate the cell lysate 3 times, 15s each time; subpackage, and store in a -80°C ultra-low temperature refrigerator for later use;

[0029] 2. Preparation of separation gel: prepare 10% separation gel solution, add TEMED to the separation gel solution, mix well and inject between two glass plates, use 7ml of glue, add water with a 200ul pipette gun to isolate the air, and make The glue surface is smooth, after standing for 20 minutes, pour off the upper layer of water, and dry it with fil...

specific Embodiment approach 3

[0037] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the proteins used for the detection of exosome purity include: TSG101, GRP78, vWF and GAPDH. Others are different from the first or second specific embodiment.

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Abstract

The invention relates to a method for detecting the milk protein and milk fat synthesis ability of cow mammary glands by using cow's milk. The purpose of the invention is to solve a problem that the milk protein and milk fat synthesis ability of mammary glands of different species and individuals of cows cannot be accurately and comprehensively reflected through existing methods for detecting the milk protein percentage and the milk fat percentage of raw milk. The method comprises the following steps: 1, preprocessing the cow's milk; 2, preparing a cow's milk exosome; 3, detecting the GlyRS content and the CAP1 content; and 4, dividing the GlyRS content by the CAP1 content to obtain the milk protein and milk fat synthesis ability of cow mammary glands. The value of the GlyRS content / the CAP1 content can reflect the milk protein and milk fat synthesis ability of the cow mammary glands, and also can solve the problem of ineffective judgment of the mammary gland's synthesis ability due to the influences of other factors on the milk protein percentage and the milk fat percentage. The method can be applied to the feed of cow raising.

Description

technical field [0001] The invention relates to a method for detecting milk protein and milk fat synthesis ability of milk cow mammary gland by using milk. Background technique [0002] Milk quality mainly refers to milk protein rate and milk fat rate, which are important economic indicators of milk. High-quality milk has higher milk protein and milk fat percentages, and has better dairy processing characteristics, so the market price is also higher. Under the same feeding and nutrition environment, the milk protein and milk fat percentages of raw milk produced by different breeds of cows and different individuals of the same breed of cows are quite different. An important indicator of differences in milk production quality between individual mammary glands. The ability of the mammary gland to synthesize milk protein and milk fat is determined by the key genes that regulate the synthesis of milk protein and milk fat in the mammary gland epithelial cells, the basic unit of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/04
CPCG01N33/04
Inventor 高学军郭思圻张明辉刘营敖金霞曲波袁肖寒
Owner NORTHEAST AGRICULTURAL UNIVERSITY