Molecular marker for detecting fatty acid character of cow milk and identification method

Abstract

The invention belongs to the technical field of biology, and discloses a molecular marker for detecting the character of fatty acid in cow milk and an identification method. The molecular marker SNP provided by the invention is g.16772853G / T, and is related to the contents of fatty acids C14: 0, C14: 1, C16: 1, C17: 0 and C18: 0 of cow milk. By utilizing the SNP provided by the invention, early screening can be realized in the dairy cow breeding process, the breeding cost can be reduced, and the fatty acid content of milk can be effectively increased or reduced according to production requirements. The method is high in detection accuracy and low in cost, and has an important application prospect in dairy cow breeding.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to molecular markers and identification methods for detecting fatty acid traits in bovine milk. Background Technology

[0002] Milk and its products are important sources of nutrition, with milk fat and its fatty acid composition playing a crucial role in nutrient supply and human health regulation. However, current technologies for the molecular detection of milk fatty acid content still suffer from limitations in methods and applications. Summary of the Invention

[0003] The technical problem to be solved by this invention is to provide molecular markers related to the fatty acid properties of milk. The technical problem to be solved is not limited to the described technical subject matter; other technical subject matter not mentioned herein will be clearly understood by those skilled in the art through the following description.

[0004] SORBS1 (sorbin and SH3 domain containing 1) participates in signaling pathways such as PPAR (ko03320) and insulin (ko04910), and has been reported as an important protein in the insulin signaling pathway in human adipose tissue, which can regulate lipid synthesis. SORBS1 Genes are associated with obesity and diabetes, and the function of the polymorphism Tyr228Ala has been reported. In the inventors' previous work on the genetic mechanism of dairy fatty acids in cows, [the following gene was identified]. SORBS1 Genes are key genes that regulate fatty acids in cow's milk.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solutions: The abbreviations in the following text represent, in order: C14:0 is tetradecanoic acid (myristic acid), C14:1 is tetradecanoic acid (or myristoleic acid), C16:1 is hexadecanoic acid (or palmitoleic acid), C17:0 is heptadecanoic acid, and C18:0 is octadecanoic acid (stearic acid).

[0006] This invention provides an application of a SNP or a substance for detecting the polymorphism or genotype of the SNP, wherein the application is one or more of the following: A1) Detect or assist in the detection of the content of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; A2) Screening or breeding of bovine breeds or strains with high or low levels of C14:0 and / or C16:1 in milk fatty acids, or low levels of one or more of C14:1, C17:0 and C18:0. A3) Screening or breeding of cattle breeds or strains with low or high levels of C14:0 and / or C16:1 in milk fatty acids, or high levels of one or more of C14:1, C17:0 and C18:0. A4) Detection or auxiliary detection of SNP polymorphisms or genotypes; A5) Cattle breeding; A6) Prepare a product for detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; A7) Prepare products from bovine breeds or varieties that have high or low contents of C14:0 and / or C16:1 in milk fatty acids, or low contents of one or more of C14:1, C17:0 and C18:0. A8) Prepare products from bovine breeds or strains that have low or high levels of one or more of C14:0 and C16:1 in milk fatty acids, or have high levels of C14:1, C17:0 and C18:0. A9) Prepare products for detecting or assisting in the detection of SNP polymorphisms or genotypes; A10) is used to prepare products for cattle breeding.

[0007] The SNP is a bovine SNP, specifically nucleotide 235 of SEQ ID NO:1 in the sequence listing, and is either G or T. The milk fatty acids of the test cattle homozygous for the SNP being T have a higher content of C14:0 or / and C16:1 than, or / and a lower content of one or more of C14:1, C17:0, and C18:0 than, the milk fatty acids of the test cattle homozygous for the SNP being G or heterozygous for the SNP being G and T.

[0008] The cattle may be bulls or cows, and the increased content of C14:0, C14:1, C16:1, C17:0 or / and C18:0 in the milk fatty acids of the bulls is reflected in the cows of their offspring.

[0009] Nucleotide 235 in SEQ ID NO:1 is the aforementioned SNP site, and its polymorphism is T / G. For ease of description, the SNP site referred to in this invention is nucleotide 235 in SEQ ID NO:1. Nucleotide 235 in SEQ ID NO:1 corresponds to nucleotide 16772853 in version number Ensembl release 80 (May 2015) or COW (UMD3.1) (URL: https: / / may2015.archive.ensembl.org / Bos_taurus / Location / View?r=26:16747442-16747442). In SEQ ID NO:1, K is G or T, and the genotype of this SNP site has the following three possibilities: TT, GT, or GG.

[0010] Those skilled in the art will understand that SEQ ID NO:1 is composed of the SNP site (nucleotide 235 of SEQ ID NO:1) and its surrounding nucleotide sequence. The amount of nucleotide sequence around the SNP site should not be used as a limiting factor for the scope of protection of this invention. It can be 25 bp, 50 bp, 70 bp, 100 bp, 150 bp, 200 bp, 300 bp, 400 bp, 500 bp, 600 bp, 700 bp, 800 bp, 900 bp, 1000 bp before and after the SNP site, or any other arbitrary value. Its function is to assist in locating the aforementioned SNP on chromosome 26 of the bovine genome.

[0011] The front and back directions mentioned in this document should be defined as front or back in a direction generally accepted by those skilled in the art, such as the 5'-3' direction.

[0012] In the above applications, the substance is any one of the following: R1) Primer composition for amplifying bovine genomic DNA fragments including the SNP; R2) PCR reagents containing the primer composition described in R1); R3) A kit containing the primer composition described in R1) or the PCR reagent described in R2).

[0013] In the above applications, the primer composition comprises any of the following: b1) A primer set consisting of the single-stranded DNA shown in SEQ ID NO:2 and the single-stranded DNA shown in SEQ ID NO:3 in the sequence listing; b2) A primer set consisting of single-stranded DNA whose nucleotide sequence is the first to second position of SEQ ID NO:1 in the sequence listing and single-stranded DNA whose nucleotide sequence is the fifth to fifth position of SEQ ID NO:1 in the sequence listing; b3) A primer set consisting of a single-stranded DNA having the same function as SEQ ID NO:2, with one or more nucleotide substitutions and / or deletions and / or additions, and a single-stranded DNA having the same function as SEQ ID NO:3, and a primer set consisting of a single-stranded DNA having one or more nucleotide substitutions and / or deletions and / or additions, and a single-stranded DNA having the same function as SEQ ID NO:3.

[0014] This invention also provides a method for detecting or assisting in the detection of components in milk fatty acids. The method includes the following steps: detecting the genotype of the aforementioned SNP in the test cow; and detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0, and C18:0 in the milk fatty acids based on the genotype. In test cows with the SNP genotype TT, the content of C14:0 and / or C16:1 in the milk fatty acids is higher than, or / and the content of one or more of C14:1, C17:0, and C18:0 is lower than, in test cows with the SNP genotype GG or GT. TT is a homozygous form of the SNP with nucleotide T. CC is a homozygous form of the SNP with nucleotide C, and GT is a heterozygous form of the SNP with nucleotides T and C.

[0015] The present invention also provides a method for cattle breeding, the method comprising any of the following: F1) The method includes detecting the genotype of the aforementioned SNP in the genome of the bovine to be tested, selecting bovines to be tested with the genotype of the SNP being TT, wherein TT is a homozygous type of the SNP being T, and the purpose of the breeding is to select bovines to be tested with high content of C14:0 or / and C16:1 or / and low content of one or more of C14:1, C17:0 and C18:0 in milk fatty acids; F2) The method includes detecting the genotype of the aforementioned SNP in the genome of the bovine to be tested, selecting bovines to be tested with the genotype of the SNP being GG, wherein GG is a homozygous type of the SNP being G, and the purpose of the breeding is to select bovines to be tested with low content of C14:0 or / and C16:1 or / and high content of one or more of C14:1, C17:0 and C18:0 in milk fatty acids.

[0016] When the method is applied to cattle breeding, if the screening target is to increase the content of milk fatty acid C14:0, cows with the TT genotype and SNP (g.16772853G / T) are selected.

[0017] When the method is applied to cattle breeding, if the screening target is to increase the content of milk fatty acid C16:1, cows with the TT genotype and SNP (g.16772853G / T) are selected.

[0018] When the method is applied to cattle breeding, if the screening target is to increase the content of milk fatty acid C14:1, cows with the SNP (g.16772853G / T) of the GG genotype are selected.

[0019] When the method is applied to cattle breeding, if the screening target is to increase the content of milk fatty acid C17:0, cows with the SNP (g.16772853G / T) of the GG genotype are selected.

[0020] When the method is applied to cattle breeding, if the screening target is to increase the content of milk fatty acid C18:0, cows with the SNP (g.16772853G / T) of the GG genotype are selected.

[0021] This invention also provides products containing substances for detecting SNP polymorphisms or genotypes in the bovine genome, including any of the following: N1) Prepare a product for detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; N2) To prepare or select products from bovine breeds or strains that have high or low levels of C14:0 and / or C16:1 in milk fatty acids. N3) Prepare products from bovine breeds or strains that are screened or selected for low or high content of C14:0 and / or C16:1 in milk fatty acids, or high content of one or more of C14:1, C17:0 and C18:0. N4) Prepare products for detecting or assisting in the detection of SNP polymorphisms or genotypes; N5) is used to prepare products for bovine breeding; The SNP is the aforementioned SNP; the content of C14:0 and / or C16:1 in the milk fatty acids of the test cows whose nucleotide of the SNP is T is higher than or / and the content of one or more of C14:1, C17:0 and C18:0 is lower than that of the test cows whose nucleotide of the SNP is G or whose nucleotide of the SNP is G and T.

[0022] In the above-mentioned products, the substance is any one of the following: R1) Primer composition for amplifying bovine genomic DNA fragments including the SNP; R2) PCR reagents containing the primer composition described in R1); R3) A kit containing the primer composition described in R1) or the PCR reagent described in R2).

[0023] The primer composition is one or more of the aforementioned b).

[0024] The present invention also provides an application, wherein the aforementioned product is used in cattle breeding.

[0025] The present invention also provides a DNA molecule comprising a DNA molecule having a strand of SEQ ID NO:1.

[0026] The present invention also provides applications of the aforementioned DNA molecules, including any of the following: Q1) Detect or assist in the detection of the content of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; Q2) Screening or breeding of bovine breeds or strains with high or low levels of C14:0 and / or C16:1 in milk fatty acids, or low levels of one or more of C14:1, C17:0 and C18:0. Q3) Screen or breed cattle breeds that have low or high levels of C14:0 and / or C16:1 or one or more of C14:1, C17:0 and C18:0 in milk fatty acids; Q4) Detect or assist in the detection of SNP polymorphisms or genotypes; Q5) Cattle breeding; Q6) Prepare a product for detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; Q7) Prepare products from bovine breeds or varieties that have high or low levels of C14:0 and / or C16:1 in milk fatty acids, or low levels of one or more of C14:1, C17:0 and C18:0. Q8) Prepare products from bovine breeds or strains that have low or high levels of one or more of C14:0 and C16:1 in milk fatty acids, or have high levels of C14:1, C17:0 and C18:0. Q9) Prepare products for detecting or assisting in the detection of SNP polymorphisms or genotypes; Q10) Preparation of bovine breeding products; The nucleotide sequence of one strand of the DNA molecule in Q) is SEQ ID NO:1; In the above text, the purpose of cattle breeding related to the SNP is to select cattle with high levels of one or more of the following fatty acids in milk: C14:0, C14:1, C16:1, C17:0, and C18:0.

[0027] In the above text, the purpose of cattle breeding related to the SNP is to select cattle with low levels of one or more of the following in milk fatty acids: C14:0, C14:1, C16:1, C17:0, and C18:0.

[0028] The cattle mentioned above can be Chinese Holstein cattle. The Chinese Holstein cattle are those defined in GB / T 3157-2023.

[0029] The purpose of this invention is to provide a molecular marker related to the fatty acid content of dairy cow milk and its application. The molecular marker g.16772853G / T is significantly or highly significantly associated with the content of milk fatty acids C14:0, C14:1, C16:1, C17:0, and C18:0. P = 0.0320 ~ P <0.0001). Attached Figure Description

[0030] Figure 1 The mutation detection result is for SNP g.16772853G / T. Detailed Implementation

[0031] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0032] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0033] In the quantitative experiments in the following examples, three replicate experiments were set up, and the average value of the results was taken.

[0034] The Chinese Holstein bull frozen semen samples were obtained from the Beijing Dairy Cattle Center. The Chinese Holstein cow samples were also obtained from the Beijing Dairy Cattle Center.

[0035] The data in the examples are all from lactation period 1. Lactation period 1 refers to the lactation period after the first delivery.

[0036] In the examples, the content of milk fatty acids was determined by gas chromatography.

[0037] The following examples used SAS 9.2 statistical software to process the data. The experimental results are expressed as least squares mean ± standard error. P<0.05 (*) indicates a significant difference. P <0.01 (**) indicates a highly significant difference.

[0038] Example 1, Related Basic Research The inventor's research group used the genotypes and milk fatty acid phenotypes of 784 Chinese Holstein cattle as data, and through genome-wide association analysis, discovered... SORBS1 The gene is a key candidate gene for milk fatty acid content (C14:1- P <2.34E-06 and C14index- P <1.09E-06). Analysis combined with functional annotations revealed... SORBS1 Genes are involved in regulating fatty acid metabolism.

[0039] Example 2: Gene Polymorphism Mining I. Forty-four Chinese Holstein bulls (from the Beijing Dairy Cattle Center) were selected as the experimental population for gene polymorphism detection in the Beijing area. The 44 Chinese Holstein bulls were randomly divided into two groups (22 bulls in each group). Genomic DNA was extracted from frozen semen samples. The DNA concentration was accurately determined using a nucleic acid quality analyzer. All DNA samples were diluted to a concentration of 50 ng / μL and mixed in equal volumes to form two DNA pools, which were then used as templates for PCR amplification.

[0040] II. According to cattle SORBS1 The gene sequence (gene ID 504625) was used to design 49 pairs of primers as shown in Table 1.

[0041] Table 1 SORBS1 Gene PCR amplification primer sequence information

[0042] Continued from Table 1

[0043] Continued from Table 1

[0044] 3. Using the pooled DNA obtained in step 1 as a template, PCR amplification was performed using each primer pair to obtain the PCR amplification products. The PCR reaction system is shown in Table 2, and the PCR reaction conditions are shown in Table 3.

[0045] Table 2. PCR reaction system

[0046] Table 3. PCR reaction conditions

[0047] Table 4 SORBS113 SNPs discovered in genes

[0048] The PCR amplification products were sequenced. The results showed that the bull population... SORBS1 A total of 13 SNPs were identified in the gene. These 13 SNPs are listed in Table 4. Among them, g.16772853G / T was identified using... SORBS1 -19F and SORBS1 The SNP site was obtained by sequencing the product from PCR amplification using primers composed of -19R (as shown in Sequence 1, where K represents G or T). The SNP site is located at position 235 from the 5' end of the PCR amplification product. g.16772853G / T is the name of the SNP. SNPs are generally named according to the pattern observed when they were first discovered. DNA has a double-stranded structure and follows the complementary base pairing principle. Sequence 1 in this invention and the sequence at the time of SNP discovery are the same double-stranded DNA. Therefore, for SEQ ID NP:1 in this invention, the polymorphic form of g.16772853G / T is G / T. Therefore, the aforementioned g.16772853G / T refers to nucleotide 235 in SEQ ID NO:1, where the polymorphic form is G / T. SEQ ID NO:1 is the reverse complementary sequence of nucleotides 16772562-16773087 on bovine chromosome 26 (link: https: / / may2015.archive.ensembl.org / Bos_taurus / Location / View?r=26:16747442-16747442). For ease of description, the SNP site referred to in this invention is nucleotide 235 in SEQ ID NO:1.

[0049] To avoid confusion, the specification still uses the G / T polymorphism method to describe g.16772853G / T.

[0050] Based on the above, we can conclude that: The SNP (g.16772853G / T) is located at position 16772853 on bovine chromosome 26. The SNP site and its surrounding nucleotides are shown in SEQ ID NO:1 of the sequence listing. Nucleotide 235 is the SNP site, corresponding to nucleotide 16772853 in Ensembl release 80 (May 2015) or COW (UMD3.1) (link: https: / / may2015.archive.ensembl.org / Bos_taurus / Location / View?r=26:16747442-16747442), and is a G / T polymorphism. In SEQ ID NO:1, K represents G or T. The genotypes for this SNP site are TT, GT, or GG. TT is a homozygous genotype with the SNP locus T, GG is a homozygous genotype with the SNP locus G, and GT is a heterozygous genotype with both the SNP locus T and G. The milk fatty acid content of cattle breeds with the TT genotype at this SNP locus is higher than that of cattle breeds with the GG or GT genotype at this SNP locus, and / or the content of one or more of C14:1, C17:0, and C18:0 is lower than that of cattle breeds with the GG or GT genotype at this SNP locus.

[0051] Example 3: Association Analysis I. Obtaining the test group The test population consisted of 1,065 Chinese Holstein cows.

[0052] II. Genotyping Each individual in the test population was genotyped separately.

[0053] 1. Take blood from the test individual and extract genomic DNA.

[0054] 2. Using genomic DNA as a template, SORBS1 -19F and SORBS1 PCR amplification was performed using primers consisting of -19R, and the PCR amplification products were then recovered and sequenced.

[0055] SORBS1 -19F refers to the primers in Table 1. SORBS1 -19F, as shown in the sequence ID NO:2 of the sequence listing; SORBS1 -19R refers to the primers in Table 1. SORBS1 -19R, as shown in sequence SEQ ID NO:3.

[0056] The reaction system for PCR amplification is shown in Table 5. The reaction conditions for PCR amplification are shown in Table 6.

[0057] The PCR amplification products of each individual were 526bp, with the 235th position being g.16772853G / T.

[0058] Table 5. PCR reaction system

[0059] Table 6. PCR Reaction Conditions

[0060] Therefore, the genotype of the SNP (i.e., g.16772853G / T) in the tested cattle can be defined according to the following rules: GG genotype: If the PCR product obtained by amplifying the genomic DNA of the pig to be tested using SORBS1-19F (SEQ ID NO:2) and SORBS1-19R (SEQ ID NO:3) contains only a DNA fragment with the nucleotide sequence of SEQ ID NO:1 and nucleotide 235 of SEQ ID NO:1 being G, and does not contain a DNA fragment with the nucleotide sequence of SEQ ID NO:1 and nucleotide 235 of SEQ ID NO:1 being T, then the aforementioned SNP genotype of the pig to be tested is GG.

[0061] TT genotype: If the PCR product obtained by amplifying the genomic DNA of the pig to be tested using SORBS1-19F (SEQ ID NO:2) and SORBS1-19R (SEQ ID NO:3) does not contain a DNA fragment with the nucleotide sequence of SEQ ID NO:1 and nucleotide 235 of SEQ ID NO:1 being G, and only contains a DNA fragment with the nucleotide sequence of SEQ ID NO:1 and nucleotide 235 of SEQ ID NO:1 being T, then the aforementioned SNP genotype of the cattle to be tested is TT.

[0062] GT genotype: If the PCR product obtained by amplifying the genomic DNA of the pig to be tested using SORBS1-19F (SEQ ID NO:2) and SORBS1-19R (SEQ ID NO:3) contains both a DNA fragment with the nucleotide sequence of SEQ ID NO:1 and nucleotide 235 of SEQ ID NO:1 being G, and a DNA fragment with the nucleotide sequence of SEQ ID NO:1 and nucleotide 235 of SEQ ID NO:1 being T, then the aforementioned SNP genotype of the pig to be tested is GT.

[0063] III. Detection of milk fatty acid content and properties Each cow in the test group underwent individual testing for milk fatty acid content.

[0064] The characteristics of milk fatty acid content include the following five indicators: C14:0, C14:1, C16:1, C17:0, and C18:0.

[0065] Each individual's record includes, in sequence, the cow's individual number, father number, mother number, grandfather number, grandmother number, maternal grandfather number, maternal grandmother number, date of birth, lactation period, calving date, and the content of C14:0, C14:1, C16:1, C17:0, and C18:0 in milk fatty acids.

[0066] IV. Association Analysis between SNP Sites and Traits Association analysis was performed on 10 indicators of milk fatty acid content traits (C14:0, C14:1, C16:1, C17:0, and C18:0) and genotypes using the MIXED procedure in SAS 9.2 software.

[0067] Association analysis uses an animal model, the specific model is as follows:

[0068] SORBS1 The association analysis results between gene g.16772853G / T and milk fatty acid content are shown in Table 7.

[0069] Table 7 SORBS1 Association analysis between gene SNP (g. 16772853G / T) and milk fatty acid content (least square mean ± standard error)

[0070] Note: ** indicates P <0.01, * indicates P <0.05, the difference is extremely significant. a,b Different superscripts in the same column of data indicate significant differences. A，B Different superscripts in the same column of data indicate highly significant differences, while the same letter superscript in the same column of data indicates no significant differences.

[0071] As shown in Table 7, g.16772853G / T was significantly or highly significantly associated with the contents of C14:0, C14:1, C16:1, C17:0, and C18:0. P = 0.0029~0.0003), for the C14:1, C17:0 and C18:0 content traits, the dominant allele is G; for the C14:0 and C16:1 content traits, the dominant allele is T.

[0072] for SORBS1The milk fatty acid C14:0 and C16:1 contents of GG genotype cows were lower than those of TT genotype cows. The milk fatty acid C14:0 and C16:1 contents of GG genotype cows were lower than those of GT genotype cows. The milk fatty acid C14:0 contents of GT genotype cows were lower than those of TT genotype cows. There was no significant difference in milk fatty acid C14:0 contents between GG genotype cows and GT genotype cows. The milk fatty acid C14:0 and C16:1 contents of GT genotype cows were not significantly different from those of TT genotype cows.

[0073] for SORBS1 The milk fatty acid C14:1, C17:0, and C18:0 contents of GG genotype cows were higher than those of TT genotype cows. The milk fatty acid C14:1, C17:0, and C18:0 contents of GG genotype cows were higher than those of GT genotype cows. The milk fatty acid C14:1, C17:0, and C18:0 contents of GT genotype cows were higher than those of TT genotype cows. The milk fatty acid C18:0 contents of GG genotype cows were not significantly different from those of GT genotype cows. The milk fatty acid C18:0 contents of GT genotype cows were not significantly different from those of TT genotype cows.

[0074] V. Analysis of Genetic Effects The significance of SNP additive effects, dominant effects, and substitution effects was tested using SAA 9.2 software.

[0075] The basic calculation formula is as follows:

[0076] The results of tests for additive effects, dominant effects, and allele substitution effects are shown in Table 8.

[0077] Table 8 SORBS1 Results of tests on additive, dominant, and substitution effects of alleles at the g.16772853G / T locus.

[0078] Note: ** express P <0.01, the difference is extremely significant.

[0079] Table 8 shows that the additive effect of g.16772853G / T on milk fatty acids C14:0, C14:1, C16:1, C17:0, and C18:0 is extremely significant; the allele substitution effect of g.16772853G / T on milk fatty acids C14:0, C14:1, C16:1, C17:0, and C18:0 is also extremely significant, meaning that each T allele substitution for a G allele leads to an increase of 0.2653 mg / 100g in C14:0. P <0.01), C14:1 decreased by 0.0962 mg / 100g ( P <0.01), C16:1 increased by 0.0801 mg / 100g ( P <0.01), C17:0 decreased by 0.0139 mg / 100g ( P <0.01) and C18:0 decreased by 0.4431 mg / 100g ( P <0.01).

[0080] The molecular markers disclosed in this invention can be used to assist in the identification of dairy cow populations with excellent milk production traits (milk fatty acid content C14:0, C14:1, C16:1, C17:0, and C18:0), and have the following advantages: simple, rapid, sensitive, reliable, stable, and accurate results, suitable for the needs of large-scale laboratory testing.

[0081] SEQ ID NO:1 (PCR product sequence): 5'--3'.

[0082] K is either G or T.

[0083] SORBS1 -19F, SEQ ID NO:2 sequence: 5'-TGGAATCAACATGTCTCACTGG-3'.

[0084] SORBS1 -19R, SEQ ID NO:3 sequence: 5'-TTTGGGACACACGTCACCTA-3'.

[0085] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. The application of a substance for detecting SNPs or their polymorphisms or genotypes, characterized in that, The application is one or more of the following: A1) Detect or assist in the detection of the content of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; A2) Screening or breeding of bovine breeds or strains with high or low levels of C14:0 and / or C16:1 in milk fatty acids, or low levels of one or more of C14:1, C17:0 and C18:

0. A3) Screening or breeding of cattle breeds or strains with low or high levels of C14:0 and / or C16:1 in milk fatty acids, or high levels of one or more of C14:1, C17:0 and C18:

0. A4) Detection or auxiliary detection of SNP polymorphisms or genotypes; A5) Cattle breeding; A6) Prepare a product for detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; A7) Prepare products from bovine breeds or varieties that have high or low contents of C14:0 and / or C16:1 in milk fatty acids, or low contents of one or more of C14:1, C17:0 and C18:

0. A8) Prepare products from bovine breeds or strains that have low or high levels of one or more of C14:0 and C16:1 in milk fatty acids, or have high levels of C14:1, C17:0 and C18:

0. A9) Prepare products for detecting or assisting in the detection of SNP polymorphisms or genotypes; A10) Products for bovine breeding; The SNP is a bovine SNP, specifically the 235th nucleotide of SEQ ID NO:1 in the sequence listing, which is either G or T.

2. The application according to claim 1, characterized in that, The substance is any one of the following: R1) Primer composition for amplifying bovine genomic DNA fragments including the SNP; R2) PCR reagents containing the primer composition described in R1); R3) A kit containing the primer composition described in R1) or the PCR reagent described in R2).

3. The application according to claim 2, characterized in that, The primer composition comprises any one of the following: b1) A primer set consisting of the single-stranded DNA shown in SEQ ID NO:2 and the single-stranded DNA shown in SEQ ID NO:3 in the sequence listing; b2) A primer set consisting of single-stranded DNA whose nucleotide sequence is the first to second position of SEQ ID NO:1 in the sequence listing and single-stranded DNA whose nucleotide sequence is the fifth to fifth position of SEQ ID NO:1 in the sequence listing; b3) A primer set consisting of a single-stranded DNA having the same function as SEQ ID NO:2, with one or more nucleotide substitutions and / or deletions and / or additions, and a single-stranded DNA having the same function as SEQ ID NO:3, and a primer set consisting of a single-stranded DNA having one or more nucleotide substitutions and / or deletions and / or additions, and a single-stranded DNA having the same function as SEQ ID NO:

3.

4. A method for detecting or assisting in the detection of the content of components in milk fatty acids, characterized in that, The method includes the following steps: detecting the genotype of the SNP described in claim 1 in the cow to be tested, and detecting or assisting in the detection of the content of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in the milk fatty acids based on the genotype.

5. A method for cattle breeding, characterized in that, The method includes any of the following: F1) The method includes detecting the genotype of the SNP in claim 1 in the genome of the bovine subject to be tested, selecting bovine subject to be tested with the genotype of the SNP being TT, wherein TT is a homozygous type of the SNP being T, and the purpose of the breeding is to select bovine subject to be tested with high content of C14:0 or / and C16:1 or / and low content of one or more of C14:1, C17:0 and C18:0 in milk fatty acids; F2) The method includes detecting the genotype of the SNP in claim 1 in the genome of the bovine subject to be tested, selecting bovine subject to be tested with the genotype of the SNP being GG, wherein GG is a homozygous type of the SNP being G, and the purpose of the breeding is to select bovine subject to be tested with low content of C14:0 or / and C16:1 or / and high content of one or more of C14:1, C17:0 and C18:0 in milk fatty acids.

6. Products containing substances that detect SNP polymorphisms or genotypes in the bovine genome, including any of the following: N1) Prepare a product for detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; N2) To prepare or select products from bovine breeds or strains that have high or low levels of C14:0 and / or C16:1 in milk fatty acids. N3) Prepare products from bovine breeds or strains that are screened or selected for low or high content of C14:0 and / or C16:1 in milk fatty acids, or high content of one or more of C14:1, C17:0 and C18:

0. N4) Prepare products for detecting or assisting in the detection of SNP polymorphisms or genotypes; N5) is used to prepare products for bovine breeding; The SNP is the SNP in claim 1.

7. The product according to claim 6, characterized in that, The substance is any one of the following: R1) Primer composition for amplifying bovine genomic DNA fragments including the SNP; R2) PCR reagents containing the primer composition described in R1); R3) A kit containing the primer composition described in R1) or the PCR reagent described in R2).

8. Application, characterized in that, The application refers to the application of the product described in claim 6 or 7 in cattle breeding.

9. A DNA molecule, characterized by, Includes a DNA molecule with a single strand of SEQ ID NO:

1.

10. Application, characterized in that, Including any of the following: Q1) Detect or assist in the detection of the content of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; Q2) Screening or breeding of bovine breeds or strains with high or low levels of C14:0 and / or C16:1 in milk fatty acids, or low levels of one or more of C14:1, C17:0 and C18:

0. Q3) Screen or breed cattle breeds that have low or high levels of C14:0 and / or C16:1 or one or more of C14:1, C17:0 and C18:0 in milk fatty acids; Q4) Detect or assist in the detection of SNP polymorphisms or genotypes; Q5) Cattle breeding; Q6) Prepare a product for detecting or assisting in the detection of one or more of C14:0, C14:1, C16:1, C17:0 and C18:0 in bovine milk fatty acids; Q7) Prepare products from bovine breeds or varieties that have high or low levels of C14:0 and / or C16:1 in milk fatty acids, or low levels of one or more of C14:1, C17:0 and C18:

0. Q8) Prepare products from bovine breeds or strains that have low or high levels of one or more of C14:0 and C16:1 in milk fatty acids, or have high levels of C14:1, C17:0 and C18:

0. Q9) Prepare products for detecting or assisting in the detection of SNP polymorphisms or genotypes; Q10) Prepare products for bovine breeding.

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