A SNP molecular marker associated with piglet size and its application

Abstract

This invention provides a SNP molecular marker associated with the piglet number trait and its application, relating to the field of biotechnology. The application includes using substances detecting SNP sites in the preparation of products predicting the piglet number trait. The SNP sites include one or more of SNP sites rs318687930 and SNP site rs329759452. These SNP sites are associated with the piglet number trait and can be used to predict this trait, as well as for screening and breeding sows with high piglet number and low birth weight with abnormalities.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a SNP molecular marker related to the piglet number trait and its application. Background Technology

[0002] The following statements are provided only as background information in relation to the present invention and do not necessarily constitute prior art.

[0003] Reproductive traits in pigs are important indicators for measuring the reproductive capacity of sows. These mainly include litter size, piglet birth weight, number of teats, milk production (litter weight at 21 days of age), and the number of piglets recovered at weaning. These traits are extremely important for pig farming. Good reproductive performance in sows means that each sow can produce more healthy piglets within a certain period, reducing breeding costs and improving production efficiency.

[0004] The litter size trait is of great significance for assessing sow reproductive efficiency and pig breeding. By studying the litter size trait, breeding pigs with high litter size potential can be screened, accelerating the transmission and fixation of superior genes, improving the reproductive capacity of pig herds, and providing a scientific basis for breeding pig selection and genetic improvement.

[0005] Therefore, the identification of genes related to the number of healthy piglets produced in pigs can provide important clues for explaining the genetic mechanisms of fetal growth and development in pigs and other mammals, and provide a theoretical basis for the genetic improvement of reproductive traits in pigs.

[0006] In view of this, the present invention is hereby proposed. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of the prior art and provide an SNP molecular marker related to the trait of healthy piglets produced and its application.

[0008] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0009] In a first aspect, a substance for detecting SNP sites is provided for use in the preparation of products that predict the number of healthy piglets produced, wherein the SNP sites include one or more of SNP sites rs318687930 and SNP sites rs329759452.

[0010] Secondly, a kit for predicting the number of healthy piglets produced in pigs is provided, the kit comprising at least one of the following primer pairs:

[0011] The primer pairs used to amplify SNP site rs318687930 have nucleotide sequences as shown in SEQ ID NO. 1 and 2, respectively; and the primer pairs used to amplify SNP site rs329759452 have nucleotide sequences as shown in SEQ ID NO. 3 and 4, respectively.

[0012] Thirdly, a method for predicting the number of healthy piglets produced in pigs is provided, the method comprising predicting sows that meet at least one of (i) and / or (ii) as sows with a high number of healthy piglets produced:

[0013] (i) The genotype of SNP locus rs318687930 is GA;

[0014] (ii) The genotype of SNP locus rs329759452 is AA.

[0015] Fourthly, a method for screening sows with a high litter size trait is provided, the method comprising screening sows with a high litter size trait predicted by the method described in the third aspect.

[0016] Fifthly, a breeding method for sows exhibiting a high litter size and high piglet count is provided, the method comprising obtaining at least one of the following genotypes in the sow:

[0017] (i) The genotype of SNP locus rs318687930 is GA;

[0018] (ii) The genotype of SNP locus rs329759452 is AA.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] This invention utilizes low-depth resequencing and genome-wide association analysis to identify SNP molecular markers associated with the piglet number trait. Two SNP loci, rs318687930 and rs329759452, were found to be associated. Sows with specific genotypes at these two loci have a higher piglet number. These SNP molecular markers can be used to predict piglet number, screen sows with high piglet number, and develop breeding methods for obtaining sows with high piglet number. The application of these SNP loci helps improve sow reproductive performance and brings greater economic benefits to the pig farming industry. Attached Figure Description

[0021] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0022] Figure 1 This is a genotype detection peak diagram obtained by capillary electrophoresis detection of SNP locus rs318687930 in an embodiment of the present invention;

[0023] Figure 2 This is a peak diagram of genotype detection obtained by capillary electrophoresis detection of SNP locus rs329759452 in an embodiment of the present invention. Detailed Implementation

[0024] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0025] cAMP response element binding protein 3 (CREB3 regulatory factor, CREBRF), also known as the Luman / CREB3 recruiting factor (LRF), is a regulator of the Luman / CREB3 protein (cyclic adenosinemonophosphate response element binding protein 3, Luman / CREB3). Studies have shown that CREBRF knockout mice have significantly lower prolactin levels than normal mice, impaired glucocorticoid signaling, and exhibit severe maternal behavioral defects, lacking the instinct to care for offspring. CREBRF gene expression is high in the ovary and uterus during estrus and postestrus, suggesting that CREBRF may be involved in cell differentiation, embryonic development, and implantation. Another study showed that CREBRF can mediate ERS pathway-mediated apoptosis of GCs and plays a crucial role in mouse follicle selection. CREBRF expression can be inhibited by E2 but induced by P4, and CREBRF can regulate decidualization during pregnancy by modulating the proliferation of mouse uterine stromal cells. Therefore, the CREBRF gene plays an important regulatory role in animal growth, development, and reproduction, especially in follicle growth and development, cell differentiation, embryonic development, and implantation. Furthermore, polymorphic changes at CREBRF gene SNP sites are correlated with estrogen levels, which in turn regulate sow reproductive efficiency. Since the number of healthy piglets born is an important indicator of sow reproductive traits, this invention investigates the correlation between CREBRF gene single nucleotide polymorphisms and the number of healthy piglets born.

[0026] This invention employs low-depth resequencing and genome-wide association analysis to analyze the association between CREBRF gene polymorphisms and important reproductive traits, and identifies SNP molecular markers in the pig genome that are significantly associated with the trait of healthy piglets produced. This invention discovered two SNP loci, rs318687930 and rs329759452, that are associated with the trait of healthy piglets produced.

[0027] The CREBRF gene SNP site rs318687930 is located at nucleotide position 51174074 on pig chromosome 16 (reference genome is Sscrofa11.1). When the genotype of this SNP site is GA, that is, nucleotide position 51174074 on pig chromosome 16 is a heterozygote of G and A, the sow has a high number of healthy piglets at first birth.

[0028] The CREBRF gene SNP site rs329759452 is located at nucleotide position 51221411 on chromosome 16 of pigs (reference genome is Sscrofa11.1). When the genotype of this SNP site is AA, the sow has a low number of primiparous malformations.

[0029] In this article, "number of healthy piglets born" refers to the number of piglets born to a female animal after parturition that are not only surviving but also healthy and free from obvious physiological defects or diseases. This indicator is an important parameter for measuring the reproductive quality of female animals and the survival rate of their offspring, especially in the livestock industry, where it is used to assess reproductive efficiency and the initial health status of young animals. "High number of healthy piglets born" refers to a number of healthy piglets born that is higher than the average number of healthy piglets born for that breed.

[0030] In this article, "birth deformity number" refers to the number of offspring with obvious morphological or structural abnormalities born during parturition. These deformities can affect any part of the offspring's body, including but not limited to the head, limbs, and internal organs, and usually lead to the offspring's inability to survive or develop normally. "Low birth deformity number" refers to a birth deformity number lower than the average birth deformity number for that breed.

[0031] Unlike the number of live piglets, the number of healthy piglets focuses more on the quality of the offspring than just their quantity. Healthy piglets have better survival ability and growth potential, which means higher breeding success rates and economic benefits for farmers. Therefore, increasing the number of healthy piglets is not just about increasing the number of piglets, but more importantly, ensuring that these piglets grow up healthily, thus providing a high-quality breeding stock for subsequent fattening or reproduction.

[0032] Based on the above findings, in a first aspect, an application is provided for a substance for detecting SNP sites in the preparation of products for predicting the number of healthy piglets produced, wherein the SNP sites include one or more of SNP sites rs318687930 and SNP sites rs329759452.

[0033] In an optional embodiment, the pig litter size trait includes at least one of primiparous healthy piglets, multiparous healthy piglets, and malformed piglets.

[0034] In an optional embodiment, the product includes a substance for detecting the SNP locus rs318687930, and the pig litter size trait includes the number of healthy piglets in a primiparous litter.

[0035] In an optional embodiment, the product includes a substance for detecting the SNP locus rs329759452, and the piglet birth rate trait includes the number of deformed piglets.

[0036] In an optional embodiment, the product includes a substance that detects SNP sites rs318687930 and rs329759452.

[0037] In optional embodiments, the substances used to detect SNP sites include, but are not limited to, one or more of the following: reagents for nucleic acid amplification, reagents for detecting nucleic acid amplification products, reagents for constructing sequencing libraries, and reagents for sequencing. More specifically, the substances used to detect SNP sites include, but are not limited to, primers, probes, enzymes for nucleic acid amplification reactions, fluorescent labels, buffer reagents, dNTPs, salts, etc. Depending on the specific detection method, those skilled in the art can select the above-mentioned reagents for nucleic acid amplification, reagents for detecting nucleic acid amplification products, reagents for constructing sequencing libraries, and reagents for sequencing according to the methods described in general and more specific textbooks, references, process manuals, product instructions, and standard documents. This invention does not impose any limitations on these selections.

[0038] In an optional embodiment, the substance for detecting SNP sites includes at least one of the following primer pairs: a primer pair for amplifying SNP site rs318687930, with nucleotide sequences as shown in SEQ ID NO. 1 and 2, respectively; and a primer pair for amplifying SNP site rs329759452, with nucleotide sequences as shown in SEQ ID NO. 3 and 4, respectively.

[0039] Secondly, a kit for predicting the number of healthy piglets produced in pigs is provided, the kit comprising at least one of the following primer pairs: a primer pair for amplifying SNP site rs318687930, the nucleotide sequences of which are shown in SEQ ID NO. 1 and 2, respectively; and a primer pair for amplifying SNP site rs329759452, the nucleotide sequences of which are shown in SEQ ID NO. 3 and 4, respectively.

[0040] Thirdly, a method for predicting the number of healthy piglets produced in pigs is provided, the method comprising predicting sows that meet (i) and / or (ii) as sows with a high number of healthy piglets produced:

[0041] (i) The genotype of SNP locus rs318687930 is GA;

[0042] (ii) The genotype of SNP locus rs329759452 is AA.

[0043] In an optional implementation, the method includes predicting sows that meet (i) and (ii) above as sows with a high litter size trait.

[0044] In an optional implementation, the method for predicting the number of healthy piglets produced in pigs includes predicting sows with a genotype of GA that meet the SNP locus rs318687930 as sows with a high number of healthy piglets produced at first litter.

[0045] In an optional implementation, the method for predicting the number of healthy piglets produced in pigs predicts sows with genotype AA that match the SNP locus rs329759452 as sows with a low number of deformed piglets.

[0046] The third aspect provides a method for predicting the number of healthy piglets produced in pigs, which includes using one or more of the SNP loci rs318687930 and SNP loci rs329759452 as the sole identification target; it can also be combined with other targets, such as other polymorphic molecular markers (e.g., SSR molecular markers, STR molecular markers or InDel molecular markers), or other screening methods known in the art to assist in the prediction of the number of healthy piglets produced in pigs.

[0047] Fourthly, a method for screening sows with a high litter size trait is provided, comprising screening sows with a high litter size trait predicted by the method described in the third aspect.

[0048] Fifthly, a breeding method is provided for sows exhibiting a high litter size and healthy piglet count trait, the breeding method comprising obtaining at least one of the following genotypes in the sow:

[0049] (i) The genotype of SNP locus rs318687930 is GA;

[0050] (ii) The genotype of SNP locus rs329759452 is AA.

[0051] The specific means by which a sow acquires a specific genotype can be implemented by those skilled in the art using conventional methods known in the art, and this invention does not limit this. In an optional embodiment, offspring with the target genotype are obtained by acquiring parents with a known genotype. In another optional embodiment, offspring with the target genotype are obtained through gene editing methods.

[0052] In an optional implementation, the pig in any of the above implementations is a Large White pig.

[0053] The present invention will be further illustrated below with specific embodiments. However, it should be understood that these embodiments are merely for the purpose of more detailed illustration and should not be construed as limiting the present invention in any way.

[0054] Example 1

[0055] I. Sample Collection

[0056] The experimental population in this embodiment consisted of 226 British Large White pigs from Rizhao Breeding Farm in Shandong, 165 Canadian Large White pigs from Xinjiang Tiankang Jiamei Breeding Branch, and 199 Danish Large White pigs from the great-grandparent breeding farm of Xinjiang Tiankang Animal Husbandry Technology Co., Ltd. A total of 590 Large White pigs were used for CREBRF gene SNP genotyping. Ear tissue samples were collected from each pig, placed in 75% alcohol, and stored at -20°C for DNA extraction.

[0057] II. Extraction and Library Construction of Pig Genomic DNA

[0058] (1) Cut an appropriate amount of pig ear tissue, cut it into pieces and put it into a 1.5mL Axgen tube.

[0059] (2) Take a 50mL BD centrifuge tube, mix proteinase K and lysis buffer to a final concentration of 0.4mg / mL evenly, and add 0.5mL lysis buffer to a 1.5mL centrifuge tube containing pig ear tissue for lysis.

[0060] (3) Place the centrifuge tubes evenly and parallel on the shaking plate of the constant temperature hybridization oven (tightly seal the tube caps to prevent liquid leakage). Incubate at 55°C for more than 6 hours (thorough mixing of the sample during digestion is very important. The criteria for judgment are the absence of obvious visible pig ear tissue and the milky consistency of the digested mixture).

[0061] (4) After the sample has been fully lysed, remove the centrifuge tubes, add 0.3 mL of saturated sodium chloride solution to each tube, mix thoroughly by inverting 6-8 times, and then place them on ice for 15 minutes.

[0062] (5) After the ice bath, centrifuge at 12,000 rpm at room temperature for 15 minutes. Carefully and slowly transfer the supernatant to a new 1.5 mL Axgen centrifuge tube (be careful to avoid pouring out the precipitate along with the supernatant, keep the pouring method consistent, and keep the amount of supernatant poured out in each tube the same).

[0063] (6) Add 0.7 mL of isopropanol (the amount of isopropanol added varies with the amount of supernatant poured out, and the two are equal in volume) to each tube, and invert until flocculent precipitate appears in the solution (if no flocculent precipitate appears, the solution can be placed in a -20°C refrigerator for 2 hours or in a 4°C refrigerator overnight).

[0064] (7) Centrifuge at 12000 rpm at room temperature for 15 minutes and remove the supernatant.

[0065] (8) Add 0.5 mL of 70% ethanol to each centrifuge tube and gently invert to thoroughly rinse the DNA that has precipitated above.

[0066] (9) Centrifuge at 10000 rpm for 30 seconds, then use a 200 μL micropipette to remove the ethanol from the centrifuge tube, leaving the precipitated DNA in the tube.

[0067] (10) Air dry DNA for 10 minutes.

[0068] (11) Pipette 0.1 mL of TE buffer into each tube to redissolve the DNA precipitate. Place the tube at 55°C for 2 hours and shake it several times to ensure that the DNA is fully dissolved.

[0069] (12) After the DNA was fully dissolved, the concentration of the extracted DNA was determined by UV spectrophotometer, and the quality of the extracted DNA was detected by agarose gel electrophoresis. The quality of the DNA was further tested using Qbuit, and qualified samples were used for library construction.

[0070] (13) Library construction: Sequencing libraries were prepared using the BGI library construction kit.

[0071] (14) Sequencing: After the library preparation is completed and the quality is checked, PE150 mode sequencing is performed using DNBSEQ-T7.

[0072] III. SNP chip genotyping and genotypic data quality control

[0073] DNA samples from 590 Large White pigs were sent to Beijing Gezhi Boya Biotechnology Co., Ltd. for 20× depth sequencing and low-depth resequencing.

[0074] Low-quality sequencing data was removed using the default parameters of fastp v0.23.2 to obtain clean reads. The clean reads were then aligned to the reference genome using the default parameters of bwa-mem v0.7.17. The alignment results were then converted into BAM files and sorted using Samtools software. Picard MarkDuplicates was used to remove duplicate reads from the sorted BAM files. Genomic variant detection was performed using GATK-HaplotypeCaller software, generating gvcf files for individual samples. GATK CombineGVCFs and GenotypeGVCFs were then used to merge and genotype the gvcf files. The filtering parameters for SNPs and Indels are shown in Table 1.

[0075] Table 1 Filtration parameters

[0076] Filtering parameters SNP Indel QD <2.0 <2.0 QUAL <30.0 <30.0 MQ <40.0 <40.0 SOR >3.0 FS >60.0 >200.0 MQRankSum <-12.5 ReadPosRankSum <-8.0 <-20.0

[0077] IV. Data Processing and Analysis

[0078] (1) Phenotypic data analysis: Phenotypic data on the number of healthy piglets produced by 590 Large White pigs were collected and organized in Excel. Descriptive statistics were performed on the phenotypic data on the number of healthy piglets produced, and the sample size, mean, standard deviation, standard error, maximum value, minimum value and quartiles of each variable were calculated.

[0079] (2) Statistical Analysis: Excel 2021 was used to calculate various polymorphism indicators. The relevant formulas for calculating polymorphism indicators are as follows:

[0080] (2.1) Gene frequency (AF)

[0081]

[0082] (2.2) Genotype frequency (GF)

[0083]

[0084]

[0085] (2.3) Genetic heterozygosity (He)

[0086]

[0087] (2.4) Effective number of alleles (Ne)

[0088]

[0089] (2.5) Polymorphic Information Content (PIC)

[0090]

[0091] (2.6) Correlation Analysis

[0092] Anova analysis was performed using the LSD method. Reproductive phenotypic characteristics of different genotypes were expressed as mean ± standard deviation. A linear model using PLINK software was used to perform association analysis between genotype and reproductive phenotype. The field effect was used as a covariate for correction. P ≤ 0.05 was considered statistically significant, and P ≤ 0.01 was considered extremely statistically significant.

[0093] V. Capillary electrophoresis detection of SNPs

[0094] (1) Determine the number of reactions N. During the test, a template-free control (nuclease-free pure water) should be tested simultaneously. N = number of samples to be tested (n) + template-free control (1) + 1.

[0095] (2) Place the enzyme mixture on ice, and thaw the remaining components at room temperature until completely melted. Mix with a vortex mixer for 10 seconds, and then centrifuge with a micro centrifuge for 10 seconds to concentrate the liquid at the bottom of the tube.

[0096] (3) Prepare the reaction system except for the template according to Table 2. Take one centrifuge tube, add each component, mix with a vortex mixer for 10s, centrifuge with a microcentrifuge for 10s, and aliquot 9μL into PCR reaction tubes. Primer sequences are shown in Table 3.

[0097] Table 2 Reaction system configuration

[0098] Serial Number Component Name Amount of standard system added (μL) Reaction number N and amount added (μL) 1 2.5×PCR reaction solution 4 4N 2 primer mixture 1 1N 3 Nuclease-free water 4 4N total 9 9N

[0099] Note: PCR products should be stored at 2–8℃ and tested within 24 hours, or stored at -20℃±5℃ and tested within 7 days. The number of freeze-thaw cycles should not exceed 3.

[0100] Table 3 Primer sequences

[0101]

[0102] (4) Add 1 μL of the sample to be tested to each of the pre-dispensed reaction tubes. Record the order of the numbers, mix using a vortex mixer for 10 s, and then centrifuge using a microcentrifuge for 10 s.

[0103] (5) Place each reaction tube in the reaction chamber of the PCR instrument and run the PCR program. The PCR amplification program is shown in Table 4.

[0104] Table 4 PCR Procedure

[0105]

[0106] (6) Electrophoresis detection of PCR products: Take an appropriate amount of PCR products and load them onto a 2% to 3% agarose gel for electrophoresis to detect whether the amplification is successful.

[0107] Capillary electrophoresis detection:

[0108] (1) The PCR amplification products were centrifuged at 3500 rpm for 2 min using a benchtop or plate centrifuge.

[0109] (2) Prepare a mixture of HiDi and Liz500. After mixing the mixture with a vortex mixer for 10 seconds, centrifuge it with a microcentrifuge for 10 seconds and dispense 9 μL into a 96-well plate. Add 1 μL of PCR product supernatant (stock solution or dilution) to each well.

[0110] (3) Cover with a rubber pad and centrifuge instantly using a benchtop or plate centrifuge;

[0111] (4) 95℃, denature for 5 min, then immediately ice bath for 3 min;

[0112] (5) Perform capillary electrophoresis using a gene analyzer. Select the "Fragment" electrophoresis method. Data analysis is performed using analytical software (Applied Biosystems).

[0113] VI. Results Analysis

[0114] Table 5

[0115]

[0116] Table 6(1)

[0117]

[0118] Table 6(2)

[0119]

[0120] Genotyping results show:

[0121] For the SNP (rs318687930) locus, 246 pigs had the GG genotype and 186 pigs had the GA genotype; for the SNP (rs329759452) locus, 246 pigs had the AA genotype and 162 pigs had the AT genotype.

[0122] Table 5 shows the allele and gene frequencies of the porcine CREBRF gene in a pig herd.

[0123] For the SNP (rs318687930) locus, the frequency of the GG genotype was 0.4331, and the frequency of the GA genotype was 0.3275. The genotype frequency of GG was higher than that of GA, and the GG allele was the dominant gene. For the SNP (rs329759452) locus, the frequency of the AA genotype was 0.4648, and the frequency of the AT genotype was 0.2852. The genotype frequency of AA was higher than that of AT, and the AA allele was the dominant gene.

[0124] Table 6 shows that at the SNP (rs318687930) locus, there were 246 GG genotypes and 186 GA genotypes in 590 Large White pigs. The CREBRF showed a highly significant difference in the number of healthy piglets in primiparous litters, with the GA genotype having a higher number than the GG genotype. At the SNP (rs329759452) locus, there were 264 AA genotypes and 162 AT genotypes in 590 Large White pigs. The CREBRF showed a highly significant difference in the number of malformed piglets in primiparous litters, with the AT genotype having a higher number than the AA genotype.

[0125] Association analysis between genotype and litter size phenotype was performed using a linear model in PLINK software, with the field effect used as a covariate for correction. The results are shown in Table 6.

[0126] The SNP (rs318687930) locus significantly affects the number of healthy piglets in a primiparous litter. The GA genotype resulted in a significantly higher number of healthy piglets in a primiparous litter than the GG genotype. Therefore, in actual pig breeding, GA genotype pigs have a higher number of healthy piglets in a primiparous litter. Capillary electrophoresis was performed to verify the SNP locus genotype; the results are shown below. Figure 1 .

[0127] The SNP (rs329759452) locus significantly affects the number of primiparous malformations. The AA genotype has a significantly lower number of primiparous malformations than the AT genotype. Therefore, in actual pig breeding, AA genotype pigs have a lower number of primiparous malformations. (See the test results below.) Figure 2 .

[0128] In summary, determining the nucleotide sequence at the SNP (rs318687930) site of the porcine CREBRF gene can help identify whether a pig has the GA or GG genotype, thus aiding in the identification of healthy piglet numbers: the GA genotype results in a significantly higher number of healthy piglets at first litter compared to the GG genotype. The GA genotype is characterized by a heterozygous G / A sequence at nucleotide position 51174074 on porcine chromosome 16; the GG genotype is characterized by a homozygous G sequence at nucleotide position 51174074 on porcine chromosome 16.

[0129] The nucleotide sequence at the SNP (rs329759452) site of the CREBRF gene in pigs can be used to determine whether an individual pig has the AT or AA genotype, thus aiding in the identification of the number of birth defects: the number of birth defects in first-time births with the AA genotype is significantly lower than that with the AT genotype. The AA genotype is a homozygous combination of A and T at position 51221411 on chromosome 16 of pigs; the AT genotype is a heterozygous combination of A and T at position 51221411 on chromosome 16 of pigs.

[0130] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of substances that detect SNP sites in the preparation of products for predicting the number of healthy piglets produced, characterized in that, The SNP locus is rs318687930, and the pig litter size trait refers to the number of healthy piglets in a primiparous litter. The number of healthy piglets in a primiparous litter is significantly higher in the GA genotype than in the GG genotype; and / or, The SNP locus is rs329759452, and the number of healthy piglets produced is the number of primiparous malformed piglets. The number of primiparous malformed piglets in the AA genotype is significantly lower than that in the AG genotype. The substance is a primer and / or a probe; The pig in question is a Large White pig.

2. A method for predicting the number of healthy piglets produced in pigs, characterized in that, Sows with the genotype GA that match the SNP locus rs318687930 are predicted to be sows with a high number of healthy piglets in their first litter. And / or, sows with genotype AA that match SNP locus rs329759452 are predicted to have a low number of birth defects; The pig in question is a Large White pig.

3. A method for screening sows with a high litter size and healthy piglet count, characterized in that, Sows with the trait of high litter size predicted by the method described in claim 2 were selected; The pig in question is a Large White pig.

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