A SNP molecular marker associated with reproductive traits in Haimen goats and its application

Abstract

This invention discloses a SNP molecular marker associated with reproductive traits in Haimen goats and its application, belonging to the fields of biotechnology and livestock breeding. The SNP molecular marker is named PRKCA g.62086257C>G. Using Haimen goat blood genomic DNA as a template, this invention determines the genotype through PCR and direct sequencing of the products. The GG and CC genotypes show a significant association with litter size in Haimen goats and can serve as effective molecular markers for improving reproductive traits. The detection method of this invention is simple, rapid, accurate, and inexpensive, which can improve the breeding efficiency of Haimen goats and has high application value.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology and livestock breeding, and relates to a SNP molecular marker related to the reproductive traits of Haimen goats and its application. Background Technology

[0002] High-quality breeds are the foundation of livestock production; therefore, improving the quality of sheep breeds through scientific means is of great significance to the development of the livestock industry.

[0003] In modern large-scale farming, the reproductive performance of ewes is a crucial factor affecting the economic benefits of farms. Reproductive traits are complex quantitative traits involving multiple processes, including ovarian follicle development, oocyte maturation, ovulation, fertilization, and embryonic development. They are regulated by multiple factors, including genetics, epigenetic modification, feeding management, hormones, and nutrition (Zhang Yanli, Guo Jiahe, Yao Xiaolei, & Wang Feng, 2022). In production, ewe reproductive traits are often improved by shortening the lambing interval or increasing the number of lambs per litter. Traditional breeding methods rely on phenotypic observation for inference and selection, which are time-consuming, labor-intensive, and often ineffective. In recent years, with the development of molecular biology, marker-assisted selection (MAS) technology has become increasingly mature. Selecting target trait genotypes using molecular markers can improve breeding efficiency and provide strong support for the breeding of goats with high fertility.

[0004] SNPs refer to genomic variations caused by a single nucleotide change. They are the simplest and most common type of heritable variation in livestock and poultry, occurring at a frequency of approximately 1 / 1000 bp throughout the genome. Due to their large number and ease of detection, SNPs have become one of the most widely used molecular markers (Lin Yuexia, Lü Yuhua, & Liao Rongrong, 2022).

[0005] Protein kinase Cα (PRKCA), also known as PKC-alpha or PKCA, encodes a protein that belongs to the protein kinase C (PKC) family, specifically the conventional PKC type. PKCs are serine-threonine kinases and can be activated by Ca2+. 2+ PRKCA is activated by diacylglycerol and is closely related to intracellular signal transduction. It is widely expressed in tissues and participates in various biological processes such as cell growth, cell adhesion, and cell volume control (Wang, 2021). PRKCA is also closely associated with reproduction; KEGG analysis revealed that PRKCA plays a role in the GnRH signaling pathway, Wnt signaling pathway, MAPK signaling pathway, and Ca2+. 2+ It is enriched in multiple reproductive-related signaling pathways, suggesting its broad involvement in reproductive regulation.

[0006] Current research on PRKCA polymorphism mainly focuses on human diseases, with a few reports on its application in the selection of superior traits in livestock. Tan Guanghui (Tan, Zhang, Li, Qin, & Wu, 2020) et al. found that two closely linked SNP sites in the 7th exon of PRKCA were significantly associated with Cherry Valley duck eggshell quality, with the CCAA type being the optimal genotype combination, which can serve as an effective molecular marker for improving duck eggshell quality. Some scholars have found that multiple PRKCA SNP sites are associated with the risk of coronary heart disease and affect glucose metabolism in coronary heart disease and lipid metabolism in ischemic stroke (Chen, 2019). In addition, PRKCA polymorphism is also associated with the risk of age-related hearing loss and asthma (Pan, 2011; Ren, 2022). However, research on the application of PRKCA polymorphism in goat reproductive traits has not yet been reported. Summary of the Invention

[0007] In order to overcome the shortcomings of existing technologies in the breeding of reproductive traits in Haimen goats, the purpose of this invention is to provide an SNP molecular marker related to the reproductive traits of Haimen goats and its application.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] In a first aspect, the present invention claims protection for the use of SNP molecular markers related to the reproductive traits of Haimen goats, or substances for detecting SNP molecular markers related to the reproductive traits of Haimen goats, in the screening and / or breeding of Haimen goats for reproductive traits:

[0010] The SNP molecular marker associated with the reproductive traits of Haimen goats is PRKCA g.62086257C>G, located at chromosome 19, 62086257bp, which is a C>G mutation.

[0011] Furthermore, the reproductive trait mentioned is the number of lambs born.

[0012] Furthermore, the screening and / or breeding of reproductive traits in Haimen goats involves using the aforementioned PRKCAg.62086257C>G molecular marker to identify the litter size trait of Haimen goats.

[0013] Furthermore, the screening and / or breeding of reproductive traits in Haimen goats also includes using the aforementioned PRKCAg.62086257C>G molecular marker to regulate the number of lambs born in Haimen goats.

[0014] Furthermore, the substance contains (or is) PCR primers for amplifying genomic DNA fragments of Haimen goats that include the SNP molecular marker.

[0015] In a specific embodiment of the present invention, the PCR primers comprise an upstream primer F1 and a downstream primer R1:

[0016] Upstream primer F1: 5'-CACCACGCTGTTTCATAG-3';

[0017] Downstream primer R1: 5'-GGCTACCCACCCAGTCA-3'.

[0018] Furthermore, using the genomic DNA of the blood of the Haimen goat to be tested as a template, PCR amplification was performed using the above-mentioned PCR primers, and the genotype of the SNP molecular marker was determined by direct sequencing of the product; individuals with homozygous GG and CC genotypes were retained, while individuals with heterozygous CG genotypes were eliminated, thereby increasing the number of lambs born in each generation of Haimen goats.

[0019] Different genotypes of PRKCA g.62086257C>G have a significant impact on the number of lambs born in the third litter of Haimen goats. The number of lambs born in the third litter of ewes with the GG genotype is significantly higher than that of ewes with the CG genotype, and the number of lambs born in the third litter of ewes with the CC genotype is significantly higher than that of ewes with the CG genotype.

[0020] In a specific embodiment of the present invention, the genotype of PRKCA g.62086257C>G is determined based on the sequencing peak diagram: when the peak diagram shows a single peak of C, the genotype is CC; when the peak diagram shows a double peak of C and G, the genotype is CG; when the peak diagram shows a single peak of G, the genotype is GG.

[0021] Secondly, this invention claims a genetic breeding improvement method for reproductive traits of Haimen goats, which uses the genomic DNA of the blood of the Haimen goat to be tested as a template, performs PCR amplification using the above-mentioned PCR primers, and determines the genotype of the SNP molecular marker by direct sequencing of the product; retains individuals with homozygous genotypes of GG and CC, eliminates individuals with heterozygous genotypes of CG, and increases the number of lambs born in each generation of Haimen goats.

[0022] Furthermore, in the above methods, different genotypes of PRKCA g.62086257C>G had a significant impact on the number of lambs born in the third litter of Haimen goats. The number of lambs born in the third litter of ewes with the GG genotype was significantly higher than that of ewes with the CG genotype, and the number of lambs born in the third litter of ewes with the CC genotype was significantly higher than that of ewes with the CG genotype.

[0023] In a specific embodiment of the present invention, the genotype is determined by direct sequencing of the product.

[0024] In a specific embodiment of the present invention, the genotype of PRKCA g.62086257C>G is determined based on the sequencing peak diagram: when the peak diagram shows a single peak of C, the genotype is CC; when the peak diagram shows a double peak of C and G, the genotype is CG; when the peak diagram shows a single peak of G, the genotype is GG.

[0025] In a specific embodiment of the present invention, the PCR amplification reaction system is 40 μL, comprising: 20 μL of 2×Taq PlusMaster Mix II; 1.6 μL each of upstream and downstream primers; 2 μL of template DNA; and 14.8 μL of deionized water. The PCR amplification reaction program is as follows: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 55℃ annealing for 20 s, and 72℃ extension for 30 s, for a total of 30 cycles; 72℃ extension for 5 min; and storage at 4℃ after PCR.

[0026] In a specific embodiment of the present invention, selecting individuals with GG and CC genotypes as parents in Haimen goat breeding can increase the number of lambs in a third litter. Individuals with the GG genotype have 1.43 more lambs in a third litter than individuals with the CG genotype, and individuals with the CC genotype have 0.56 more lambs in a third litter than individuals with the CG genotype.

[0027] The aforementioned SNP molecular markers related to the reproductive traits of Haimen goats also fall within the scope of protection of this invention.

[0028] The substances described above used for detecting SNP molecular markers related to the reproductive traits of Haimen goats are also within the scope of protection of this invention.

[0029] Furthermore, the substance is a PCR primer for amplifying a Haimen goat genomic DNA fragment containing the SNP molecular marker, or a kit containing the PCR primer. The specific nucleotide sequence of the PCR primer is as described above.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] This invention marks the first discovery of an effective SNP molecular marker in the PRKCA gene that influences the litter size of Haimen goats. Using the kit provided by this invention, the genotype of the molecular marker is detected by PCR amplification and direct sequencing of the product. This method is simple, rapid, accurate, and inexpensive. Screening Haimen goats using the molecular markers of this invention, retaining individuals with the GG and CC genotypes for genetic selection while eliminating individuals with the CG genotype, can improve the reproductive performance of Haimen goats, accelerate the breeding progress of Haimen goats, and has high application value. Attached Figure Description

[0032] Figure 1 The results of 2% agarose gel electrophoresis of the mixed-pool PCR amplification product of PRKCA g.62086257C>G site (primer P1) are shown, where M represents DL2000 DNA Marker and 1 represents PRKCA g.62086257C>G.

[0033] Figure 2This is a sequencing image of the PCR amplification product of the PRKCA g.62086257C>G molecular marker (primer P1); the arrows indicate the mutation sites.

[0034] Figure 3 The correlation diagram is shown between PRKCA g.62086257C>G and the number of lambs born in the third litter of Haimen goats. Detailed Implementation

[0035] The technical solution of the present invention will be further described in detail below with reference to the embodiments and related drawings. However, it is worth noting that the embodiments of the present invention are not limited thereto, and those skilled in the art can implement them in other different forms without departing from the spirit and purpose of the present invention.

[0036] Example 1: Genotyping and Identification of SNP Molecular Markers in the PRKCA Gene of Haimen Goat

[0037] 1. Collection of experimental animals and samples

[0038] The Haimen goats used in this experiment were sourced from Jiangsu Jinsheng Goat Breeding Technology Development Co., Ltd., totaling 1100 individuals. All goats were healthy and raised under identical conditions and environments. 10 mL of blood was collected from the jugular vein of each pregnant ewe, placed in an anticoagulant tube containing EDTA, and stored at -20°C. Phenotypic information, such as the number of lambs born, was collected for subsequent association analysis experiments.

[0039] 2. Main Instruments

[0040] Pipettes (Eppendorf), electronic balance (HENGJI), microwave oven (Galanz), refrigerator (Haier), handheld centrifuge (SCILOGEX, S1010E), vortex mixer (Dalong), digital display constant temperature water bath (Changzhou Putian, HH-G2), high-speed refrigerated centrifuge (Eppendorf, 5424R), micro spectrophotometer (NANODROP2000), PCR instrument (Applied Biosystems), electrophoresis apparatus (Beijing Liuyi, DYY-6C), fully automated digital gel imaging system (Tanon, Tanon-4100).

[0041] 3. Main reagents

[0042] TIANGEN Blood Genomic DNA Extraction Kit (Centrifuge Column), 50×TAE (Solarbio), Agarose (BIOWEST), 2×Taq Plus Master MixII (Dye Plus) (Vazyme, P213-03), 10000×TS-GelRed Nucleic Acid Gel Dye (TSINGKE, TSJ003), DL2000 DNA Marker (TSINGKE, TSJ011).

[0043] 4. Methods

[0044] 4.1 Extraction of genomic DNA from goat blood

[0045] Five hours in advance, remove the goat blood from the -20°C freezer and place it in a 4°C freezer to thaw. Once the blood is completely thawed, extract DNA from the whole blood according to the instructions of the TIANGEN Blood Genomic DNA Extraction Kit.

[0046] DNA concentration and quality were detected using a micro-spectrophotometer, including OD. 260 / 280 It should be between 1.80 and 2.00, OD 260 / 230 The value should be between 1.80 and 2.20. DNA samples that pass the test should be stored at -20°C.

[0047] 4.2 Primer Design for Candidate SNP Sites

[0048] Information on SNP sites related to the PRKCA gene was found using the Ensembl database (https: / / asia.ensembl.org / index.html), and genotyping primers were designed using Primer Premier 5 software. SNP site information and primer information are shown in Tables 1 and 2.

[0049] Table 1. SNP locus information of PRKCA gene

[0050]

[0051] Table 2 PRKCA SNP genotyping primer sequences

[0052]

[0053] 4.3 Candidate SNP site polymorphism and primer verification

[0054] 4.3.1 Mixing Pool Preparation

[0055] Thirty DNA samples were randomly selected from 1100 samples from Haimen goats. 1 μL of each sample was added to the same 1.5 mL centrifuge tube to create a DNA pool for subsequent verification of candidate SNP polymorphisms and primers. Another 30 DNA samples were randomly selected, and the above procedure was repeated to create two DNA pools.

[0056] 4.3.2 Mixed-pool PCR amplification

[0057] (1) PCR amplification system (20 μL): 10 μL of 2×Taq Plus Master Mix II; 0.8 μL each of forward and reverse primers; 1 μL of template DNA; 7.4 μL of deionized water.

[0058] (2) PCR amplification program: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 55℃ annealing for 20 s, 72℃ extension for 30 s, for a total of 30 cycles; 72℃ extension for 5 min; after PCR, store at 4℃.

[0059] 4.3.3 Agarose gel electrophoresis of mixed-cell PCR products

[0060] (1) Preparation of 2% agarose gel: Measure 1g of agarose and 50mL of 1×TAE and pour them into an Erlenmeyer flask. Heat in a microwave oven on high for 3 minutes until the solution is clear and transparent; otherwise, extend the heating time appropriately. After standing until the solution is no longer hot to the touch, add 5μL of nucleic acid dye, mix thoroughly, pour onto a plate, and wait for solidification.

[0061] (2) After the agarose gel solidifies, the sample loading operation is performed. The PCR product loading volume is 6 μL and the DNA Marker loading volume is 4 μL. Gently place the agarose gel into the electrophoresis tank containing 1×TAE (the 1×TAE liquid level should cover the gel surface), and electrophoresis at 140V for 35 min.

[0062] (3) After electrophoresis, the gel was transferred to a fully automated digital gel imaging system for observation.

[0063] The mixed-pool DNA was subjected to agarose gel electrophoresis, and the results are as follows: Figure 1 As shown, the PRKCA SNP site shows a clear band at 432 bp, consistent with the expected fragment size, and without any nonspecific bands, indicating good primer specificity and allowing for subsequent operations.

[0064] 4.4 Genotyping

[0065] 4.4.1 Sample PCR amplification

[0066] (1) PCR amplification system (40μL): 20μL of 2×Taq Plus Master Mix II; 1.6μL each of upstream and downstream primers; 2μL of template DNA; 14.8μL of deionized water.

[0067] (2) PCR amplification program: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 55℃ annealing for 20 s, 72℃ extension for 30 s, for a total of 30 cycles; 72℃ extension for 5 min; after PCR, store at 4℃.

[0068] 4.4.2 Sequencing of Sample PCR Products and Determination of Results

[0069] The PCR products and primer P1 were sent to Qingke Biotechnology for sequencing. The sequencing results were compared and analyzed using SnapGene software, and four SNP mutation sites were identified: PRKCA g.62086257C>G, PRKCA g.62086221G>A, PRKCA g.62086421C>T, and PRKCA g.62086425A>G.

[0070] Determine the genotype of the PRKCA SNP locus based on the sequencing peak diagram:

[0071] (1) PRKCA g.62086257C>G site: When the peak diagram shows a single peak of C, the genotype is CC; when the peak diagram shows a double peak of C and G, the genotype is CG; when the peak diagram shows a single peak of G, the genotype is GG. Figure 2 ).

[0072] (2) PRKCA g.62086221G>A site: When the peak diagram shows a single peak G, the genotype is GG; when the peak diagram shows a double peak of G and A, the genotype is AG; when the peak diagram shows a single peak A, the genotype is AA.

[0073] (3) PRKCA g.62086421C>T site: When the peak diagram shows a single peak C, the genotype is CC; when the peak diagram shows a double peak of C and T, the genotype is CT; when the peak diagram shows a single peak T, the genotype is TT.

[0074] (4) PRKCA g.62086425A>G site: When the peak diagram shows a single peak of A, the genotype is AA; when the peak diagram shows a double peak of A and G, the genotype is AG; when the peak diagram shows a single peak of G, the genotype is GG.

[0075] The above results indicate that PCR amplification of Haimen goat genomic DNA using primer P1, followed by genotyping via direct sequencing, enables rapid and accurate identification of the SNP loci of the Haimen goat PRKCA gene.

[0076] Example 2: Statistical analysis of PRKCA gene SNP polymorphism and its relationship with the high fertility trait of Haimen goats.

[0077] 1. Statistics of population genetic parameters of PRKCA SNP loci

[0078] Genotyping was performed on 1100 Haimen goats using the primers and method designed in Example 1, and the population genetic parameters of the two loci were calculated, including genotype frequency, allele frequency, homozygosity, heterozygosity, effective number of alleles, polymorphism information content, and Hardy-Weinberg equilibrium P-value. The results are shown in Table 3.

[0079] (1) PRKCA g.62086257C>G has three genotypes CC, CG and GG in Haimen goats, among which CC is the dominant genotype and C is the dominant allele. This locus shows low polymorphism (PIC<0.25) and is in Hardy-Weinberg disequilibrium (P<0.05).

[0080] (2) PRKCA g.62086221G>A has three genotypes AA, AG and GG in Haimen goats, among which GG is the dominant genotype and G is the dominant allele. This locus shows low polymorphism (PIC<0.25) and is in Hardy-Weinberg equilibrium (P>0.05).

[0081] (3) PRKCA g.62086421C>T has three genotypes CC, CT and TT in Haimen goats, among which CT is the dominant genotype and T is the dominant allele. This locus shows moderate polymorphism (0.25<PIC<0.5) and is in Hardy-Weinberg equilibrium (P>0.05).

[0082] (4) PRKCA g.62086425A>G has three genotypes AA, AG and GG in Haimen goats, among which AG is the dominant genotype and G is the dominant allele. This locus shows moderate polymorphism (0.25<PIC<0.5) and is in Hardy-Weinberg equilibrium (P>0.05).

[0083] Wherein, PRKCA g.62086421C>T and PRKCA g.62086425A>G are in linkage, and their population genetic parameters are identical.

[0084] Table 3 Population genetic parameters of PRKCA gene

[0085]

[0086]

[0087] 2. Association between PRKCA SNP loci and high prolificacy traits of Haimen goats

[0088] Using SAS (8.0) software and the GLM program, least squares statistical analysis was performed to analyze the association between different genotypes of the four SNP loci of the PRKCA gene and the number of lambs born in Haimen goats.

[0089] Using genotype and season as fixed effects, the model is: Y ijk =μ+G i +S j +e ijk In the formula, Y ijk G represents the phenotypic value of the number of lambs born to an individual; μ represents the population mean; G represents the population mean. i Genotype effect; S j This is due to seasonal effects; e ijk This is random error.

[0090] 2.1 Association analysis between PRKCA SNP loci and lambing number in Haimen goats

[0091] Fixed effects other than genotype effects that were not significantly associated with individual phenotypic values ​​were removed. Association analysis was performed between PRKCA gene SNP loci and the number of lambs born in Haimen goats. Results are expressed as "least square mean ± standard error".

[0092] The results are shown in Table 4 and Figure 3 As shown, the PRKCA g.62086257C>G locus was significantly associated with the number of lambs born in the third litter in Haimen goats. Ewes with the GG genotype had a significantly higher number of lambs born in the third litter than those with the CG genotype, and ewes with the CC genotype had a significantly higher number of lambs born in the third litter than those with the CG genotype. The remaining loci were not significantly associated with the number of lambs born.

[0093] Table 4. Association between PRKCA gene SNP sites and lambing number in Haimen goats.

[0094]

[0095] Note: Different lowercase letters in the same column indicate significant differences (P < 0.05).

[0096] Example 3: Method for improving the reproductive traits of Haimen goats

[0097] The main methods for improving the reproductive traits of Haimen goats include the following steps:

[0098] 1. Collect blood samples from the Haimen goats to be tested and extract genomic DNA from the blood.

[0099] 2. Using primer P1 as described in Example 1, PCR amplification was performed with blood genomic DNA as a template, and the genotype was determined by direct sequencing of the product.

[0100] in:

[0101] (1) PCR amplification system (40μL): 20μL of 2×Taq Plus Master Mix II; 1.6μL each of upstream and downstream primers; 2μL of template DNA; 14.8μL of deionized water.

[0102] (2) PCR amplification program: 95℃ pre-denaturation for 3 min; 95℃ denaturation for 15 s, 55℃ annealing for 20 s, 72℃ extension for 30 s, for a total of 30 cycles; 72℃ extension for 5 min; after PCR, store at 4℃.

[0103] (3) Sequencing: The PCR product and primer P1 were sent to Qingke Biotechnology Co., Ltd. for direct sequencing.

[0104] (4) Genotype determination: The sequencing peak diagrams of the PRKCA g.62086257C>G site were compared and analyzed using SnapGene software. When the peak diagram showed a single peak of C, the genotype was CC; when the peak diagram showed a double peak of C and G, the genotype was CG; and when the peak diagram showed a single peak of G, the genotype was GG.

[0105] 3. The GG and CC genotypes of PRKCA g.62086257C>G can serve as effective molecular markers for improving the reproductive traits of Haimen goats.

[0106] Selecting individuals with the GG and CC genotypes as parents in Haimen goat breeding can increase the number of lambs born in the third litter of ewes. Individuals with the GG genotype have 1.43 more lambs born in the third litter than individuals with the CG genotype, and individuals with the CC genotype have 0.56 more lambs born in the third litter than individuals with the CG genotype.

[0107] By using the SNP molecular markers of this invention to screen Haimen goats, retaining individuals with the GG and CC genotypes for genetic selection and eliminating individuals with the CG genotype, the breeding goal of improving the reproductive performance of Haimen goats can be achieved.

Claims

1. Application of PCR primers for detecting SNP molecular markers associated with reproductive traits in Haimen goats in the screening of reproductive traits or breeding of Haimen goats with high reproductive traits: The SNP molecular markers associated with the reproductive traits of Haimen goats are: PRKCA g.62086257 C>G, located at 62086257 bp on chromosome 19, is a C to G mutation; the reproductive trait described is the number of lambs born; PRKCA g.62086257 C>G Different genotypes have a significant effect on the number of lambs born in the third litter of Haimen goats. The number of lambs born in the third litter of ewes with the GG genotype is significantly higher than that of ewes with the CG genotype, and the number of lambs born in the third litter of ewes with the CC genotype is significantly higher than that of ewes with the CG genotype. The PCR primers comprise an upstream primer F1 and a downstream primer R1: Upstream primer F1: 5'-CACCACGCTGTTTCATAG -3'; Downstream primer R1: 5'-GGCTACCCACCCAGTCA-3'.

2. The application according to claim 1, characterized in that, Using the genomic DNA of the blood of the Haimen goat to be tested as a template, PCR amplification was performed using the PCR primers described above, and the genotype of the SNP molecular marker described in claim 1 was determined by direct sequencing of the product; individuals with homozygous genotypes of GG and CC were retained, while individuals with heterozygous genotypes of CG were eliminated, thereby increasing the number of lambs born in each generation of Haimen goats.

3. The application according to claim 2, characterized in that, Based on the sequencing peak diagram PRKCA Determination of the C>G genotype for g.62086257: When the peak diagram shows a single peak of C, the genotype is CC; when the peak diagram shows a double peak of C and G, the genotype is CG; when the peak diagram shows a single peak of G, the genotype is GG.

4. A method for genetic breeding improvement of reproductive traits in Haimen goats, characterized in that: The reproductive trait mentioned is the number of lambs born; using the genomic DNA of the blood of the Haimen goat to be tested as a template, PCR amplification is performed using the PCR primers described in claim 1, and the genotype of the SNP molecular marker described in claim 1 is determined by direct sequencing of the product; individuals with homozygous genotypes of GG and CC are retained, while individuals with heterozygous genotypes of CG are eliminated, thereby increasing the number of lambs born in each generation of Haimen goats; PRKCA The different genotypes of g.62086257 C>G have a significant impact on the number of lambs born in the third litter of Haimen goats. The number of lambs born in the third litter of ewes with the GG genotype is significantly higher than that of ewes with the CG genotype, and the number of lambs born in the third litter of ewes with the CC genotype is significantly higher than that of ewes with the CG genotype.

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