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Method for identifying and breeding A2-type milk-producing livestock and method for producing A2-type milk by using A2-type milk-producing livestock

A livestock and milk production technology, applied in the field of biological genetics and breeding, can solve the problems of complicated detection process, complicated operation and low throughput, etc.

Pending Publication Date: 2020-04-28
BIOYONG TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method requires pre-processing of specific amino acid sequences, and repeated optimization of HPLC parameters and ESI-QqQ conditions. The detection process is cumbersome and complicated, and it cannot meet the development needs of the modern dairy industry.
[0009] In summary, the detection methods for A1β-casein and A2β-casein generally have the defects of cumbersome operation and low throughput.

Method used

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  • Method for identifying and breeding A2-type milk-producing livestock and method for producing A2-type milk by using A2-type milk-producing livestock
  • Method for identifying and breeding A2-type milk-producing livestock and method for producing A2-type milk by using A2-type milk-producing livestock
  • Method for identifying and breeding A2-type milk-producing livestock and method for producing A2-type milk by using A2-type milk-producing livestock

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1. Test result reproducibility verification

[0090] 1. Sample pretreatment

[0091] a) Commercially available ordinary milk (commercial name: Sanyuan pure milk; manufacturer: Beijing Sanyuan Food Co., Ltd.; place of origin: China) and commercially available A2 milk (commercial name: extreme A2-β casein fresh milk; produced Supplier: Beijing Sanyuan Food Co., Ltd.; place of origin: China), mix the two milks evenly at a volume ratio of 1:1.

[0092] b) Accurately draw 18 identical mixed milk samples, 10 μl each, add 920 μl sodium bicarbonate buffer solution to each milk sample, and vortex for 30 seconds at 1200 rpm on a vortex mixer.

[0093] c) Add 10 μl of dithiothreitol solution, and react at a constant temperature of 50° C. for 30 minutes.

[0094] d) After cooling to room temperature, 30 μl of iodoacetamide solution was added, and stood at room temperature in the dark for 30 minutes.

[0095] e) Add 10 μl of trypsin solution, and perform enzymatic hydroly...

Embodiment 2A2

[0108] Embodiment 2A2-beta casein semi-quantitative detection

[0109] 1. Sample pretreatment

[0110] a) Select the three-yuan pure milk sample (trade name: three-yuan pure milk; manufacturer: Beijing Sanyuan Food Co., Ltd.; place of origin: China) as the A1 / A2 hybrid milk in this embodiment;

[0111] b) Select A2 full-fat milk powder sample (trade name: A2 full-fat adult milk powder; manufacturer: A2 Co., Ltd.; place of origin: New Zealand; importer: China National Land Reclamation Holdings Shanghai Company) as the A2 homozygous milk sample in this example ;

[0112] c) Accurately weigh 1.00 g of A2 whole milk powder sample, add purified water, and vortex on a vortex mixer at 3000 rpm for 2 minutes to completely dissolve the milk powder;

[0113] d) Mix A1 / A2 heterozygous milk and A2 homozygous milk according to the volumes in the table below;

[0114] e) Table 3. Dairy Mixing Volume Table

[0115]

[0116]

[0117] f) Accurately draw 10 μl of the mixed milk sample...

Embodiment 3

[0132] Example 3 commercially available pure milk detection

[0133] 1. Sample pretreatment

[0134] a) Accurately draw 10 μl of pure milk sample (trade name: Sanyuan pure milk; manufacturer: Beijing Sanyuan Food Co., Ltd.; place of origin: China), add 920 μl of sodium bicarbonate buffer, and vortex on a vortex mixer at 1200rpm for 30s .

[0135] b) Add 10 μl of dithiothreitol solution, and react at a constant temperature of 50° C. for 30 minutes.

[0136] c) After cooling to room temperature, 30 μl of iodoacetamide solution was added, and left standing at room temperature in the dark for 30 minutes.

[0137] d) Add 10 μl of trypsin solution, and perform enzymatic hydrolysis at a constant temperature of 37° C. for more than 2 hours.

[0138] e) Add 20 μl of pure formic acid to terminate the reaction. Obtain the sample treatment solution.

[0139] f) Aspirate 20 μl of sample treatment solution and mix with 20 μl of SA matrix, and vortex at 1200 rpm for 30 seconds on a vort...

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Abstract

The invention discloses a method for identifying and breeding A2 type milk-producing livestock. According to the method, A2 type milk-producing livestock is identified through standard characteristicpolypeptide groups of A1 beta-casein and A2 beta-casein, and thus breeding can be completed. The milk-producing livestock comprises milk-producing cows and sheep, and the standard characteristic polypeptide sequences of the A1 beta-casein and the A2 beta-casein are respectively as shown in SEQ ID No.1 and SEQ ID No.2. The invention also discloses a method for producing A2-type milk by using the method.

Description

technical field [0001] The invention belongs to the field of biological genetic breeding, and in particular relates to a method for identifying and breeding A2-type milk-producing livestock using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF MS) and a method for producing A2-type milk. [0002] technical background [0003] Casein is the protein with the highest content in dairy products, accounting for about 80% of milk protein content. According to the amino acid composition and electrophoretic behavior, casein can be divided into αs-casein, β-casein and κ-casein, of which β-casein accounts for about 30-35% of the total bovine casein. There are many variants of β-casein, the most common ones are A1β-casein and A2β-casein, the difference between the two is that there is a variation in the 67th position of the amino acid chain, the former is histidine, and the latter is proline acid. Dairy products containing both A1β-casein and A2β...

Claims

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Application Information

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IPC IPC(8): G01N27/62G01N33/68G01N27/626
CPCG01N27/628G01N33/6851G01N2560/00
Inventor 陈莲莲吕倩何昆付书辉马庆伟向华
Owner BIOYONG TECH
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