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A method for screening dairy cows with different lactoferrin content and the kit used therefor

A lactoferrin and kit technology, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc., can solve the problem of high price of natural lactoferrin

Active Publication Date: 2022-02-25
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the extremely low content of natural lactoferrin in milk, the price of natural lactoferrin as a food additive and nutritional fortifier is still relatively expensive

Method used

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  • A method for screening dairy cows with different lactoferrin content and the kit used therefor
  • A method for screening dairy cows with different lactoferrin content and the kit used therefor
  • A method for screening dairy cows with different lactoferrin content and the kit used therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1, Discovery of T53538908ASNP and classification of cow genotypes based on the SNP locus

[0065] Considering the influence of feeding environment factors on individuals, dairy cows from the same cattle farm (specifically Treasure Island Cattle Farm in the southern suburbs of Beijing) were selected for the experiment. At the same time, in order to control the error caused by the differences among individuals, individuals with the same number of lactation days (specifically the 15th day of lactation), 2 or 3 parities, and close sampling time were selected to ensure that the sampling time and individual differences were reduced as much as possible. to the minimum. Finally, according to the milk production record of the individual's previous parity, the individual's 305-day milk production, milk fat amount and milk protein amount are counted; the individual dairy cows with a 305-day milk output greater than 12,000 kg, a milk fat amount greater than 450 kg, and a mi...

Embodiment 2

[0089] Example 2. Establishment of a genotyping method based on T53538908A SNP for dairy cows to be tested

[0090] 1. Preparation of primer pairs

[0091] Primer pair 1, primer pair 2, primer pair 3, primer pair 4, and primer pair 5 were designed to amplify the target sequence including the T53538908A SNP.

[0092] The nucleotide sequences and amplified fragment sizes of the upstream and downstream primers of each primer pair are shown in Table 4.

[0093] Table 4

[0094]

[0095] 2. Establishment of genotyping method based on T53538908A SNP for dairy cows to be tested

[0096] 1. Using the genomic DNA of the dairy cow to be tested as a template, a primer pair (primer pair 1, primer pair 2, primer pair 3, primer pair 4 or primer pair 5) is used for PCR amplification to obtain a PCR amplification product.

[0097] 2. After step 1 is completed, the PCR amplification product is detected by 1% (m / v) agarose gel electrophoresis.

[0098] For some test results, see figure 1 ...

Embodiment 3

[0103] Example 3. Evaluating the content of lactoferrin in milk by detecting the genotype of the cow to be tested based on the T53538908A SNP

[0104] Among the kits for detecting the content of lactoferrin in milk, the results based on the principle of elisa enzyme-linked immunosorbent reaction have low repeatability, and there are only 1-2 kinds in China; foreign kits are generally not in stock, and the waiting period is 3-4 months , The storage time of test samples will affect the test results. The LTF gene encodes lactoferrin, and the detection of the expression level of the LTF gene is simple, accurate and stable. Therefore, the content of lactoferrin can be evaluated by detecting the normalized expression of LTF gene. The higher the normalized expression level of the LTF gene, the higher the lactoferrin content.

[0105] 1. Evaluate the content of lactoferrin in the milk of 6 dairy cows in Tinyindao Cattle Farm in the southern suburbs of Beijing by detecting the genoty...

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Abstract

The invention discloses a method for screening cows with different lactoferrin contents and a kit used therefor. The method is as follows: detecting whether the genotype of the dairy cow to be tested is genotype I, genotype II or genotype III, the lactoferrin content in the milk of the cow of genotype I is less than that of the cow of genotype II, and the content of lactoferrin in the milk of the cow of genotype II is Cows with lower lactoferrin content than genotype III, cows with genotype I, cows with genotype II and cows with genotype III are TT homozygous, TA heterozygous and AA homozygous respectively based on the T53538908A SNP , T53538908ASNP is the 53538908th nucleotide of the 22nd chromosome of the cow genome. The method provided by the invention can screen individual dairy cows with higher lactoferrin content in milk and has higher accuracy. The invention has great application value for establishing high-quality dairy cow population.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for screening cows with different lactoferrin contents and a kit used therefor. Background technique [0002] Lactoferrin encoded by LTF gene (Genebank No. 280846) is a naturally active non-heme iron-binding glycoprotein in milk. It is mainly expressed and secreted by mammary epithelial cells and is one of the important nutrients in milk. Lactoferrin has a wide range of biological activities, participates in iron transport, plays an important role in the innate immune system, and has been shown to have inhibitory effects on certain cancer cells. Therefore, lactoferrin is a food additive with great development potential. Lactoferrin is an important nutritional ingredient required by infants and has been widely used in infant formula milk powder. Studies have shown that lactoferrin can reduce the incidence of diarrhea and respiratory tract infection in infants an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 姜力陈一丹杜成祥张昱张勤
Owner CHINA AGRI UNIV
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