SNP (Single Nucleotide Polymorphism) marker for increasing goat weight and application thereof

Abstract

The invention discloses an SNP (Single Nucleotide Polymorphism) marker related to goat weight and application thereof. The marker is located on an exon of an FAM184B gene of a goat chromosome 6, the specific SNP marker is T / C base mutation at the 426bp position of SEQ ID NO: 1 in a sequence table, and the weight of TT and TC genotypes is significantly higher than that of a CC genotype individual. According to the present invention, the Musheng black goat is adopted as the research object, the DNA sequence of the goat FAM184B gene is subjected to PCR amplification, and the influence of the mutation site polymorphism on the goat body weight is analyzed, such that the method can be used for breeding the Musheng black goat, and the breeding of the Musheng black goat variety using the Musheng black goat as the breeding material is accelerated; and a basic marker resource is provided for marker-assisted selective breeding of goat weight traits.

Description

Technical Field

[0001] The invention belongs to the technical field of animal molecular breeding, and relates to the application of SNP markers related to goat weight traits in the breeding of Macheng black goats. Background Art

[0002] The purpose of meat livestock production is to produce meat. The heavier the individual, the more meat it produces. Therefore, weight is a key indicator for evaluating goat productivity and economic benefits. The phenotype of traits is controlled by genes. Therefore, molecular marker-assisted selection based on the genes that control traits has the advantages of simple operation and early selection, which can effectively shorten the breeding period and improve breeding efficiency.

[0003] A large number of omics analyses have shown that the family with sequences similarity 184 member B (FAM184B) gene is associated with the body shape and weight traits of livestock. Based on GWAS analysis, it has been determined that this gene affects the body shape of horses, the height of donkeys, the 14-month-old weight and muscle growth traits of sheep, and the weight of chickens and pigeons. It was found that one SNP of this gene affects the body length and tarsal circumference of horses, and seven SNPs are significantly associated with carcass weight. In summary, the FAM184B gene is closely related to the growth traits of livestock and poultry, and is related to muscle growth and bone development.

[0004] Macheng Black Goat is an excellent black goat meat breed in the Dabie Mountains. It has the growth characteristics of fast growth rate and good fattening ability. Based on the whole genome selection signal analysis, Macheng Black Goat was divided into high body weight group and low body weight group, and the positive selection signal was detected. It was found that a positively selected SNP marker of the FAM184B gene affected body weight, providing a new marker resource for the molecular marker-assisted selection breeding of weight traits of Macheng Black Goat. Summary of the invention

[0005] The invention aims to provide a technical method for goat weight breeding.

[0006] According to an embodiment of the present invention, the SNP marker is located at the 426th base of the nucleotide sequence shown in SEQ ID NO: 1, and the weight of individuals with TT and TC genotypes of the SNP marker is significantly higher than that of individuals with CC genotypes. By detecting the above SNP marker of goats, the weight of goats can be evaluated according to the genotype of the SNP marker. Therefore, the SNP marker of the present invention is closely related to the weight of goats, and can be effectively used for molecular marker-assisted breeding of goats, and then goat breeding materials can be selected according to actual breeding needs, so that excellent goat varieties can be accurately and efficiently bred, and the efficiency and accuracy of breeding can be improved.

[0007] The present invention provides a primer pair for detecting a SNP marker as claimed in claim 1. According to an embodiment of the present invention, the primer has a nucleotide sequence shown in SEQ ID NO: 2 and SEQ ID NO: 3. The primer pair of the present invention can effectively perform PCR amplification on the fragment where the SNP marker related to the weight of the goat to be tested is located, and the detection of the SNP marker can be effectively achieved by sequencing, and the genotype of the SNP marker of the goat to be tested can be determined, and then the weight of the goat to be tested can be effectively predicted. Specifically, the weight of individuals with TT and TC genotypes of the SNP site is significantly higher than that of individuals with CC genotype, which can be used as an important criterion for judging the weight of goats. In the goat breeding work, individuals with TT genotype of the SNP site can be retained for breeding, and individuals with CC genotype of the SNP marker site can be eliminated. It is also possible to breed CT genotype individuals with TT genotype individuals to obtain more TT genotype offspring, so as to achieve low-cost and high-accuracy breeding of excellent goat varieties, thereby gradually improving the weight of the goat population.

[0008] The present invention has the following beneficial effects: (1) The SNP marker provided by the present invention is significantly correlated with the weight of Macheng black goats, and the weight of Macheng black goats with TT and TC genotypes is significantly higher than that of Macheng black goats with CC genotypes; (2) The SNP marker can be used for auxiliary selection of the weight of Macheng black goats to screen out Macheng black goats with high weight, which has important practical application value for further improving the weight of Macheng black goats and using Macheng black goats as materials for variety (or strain) breeding. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] The above aspects of the present invention will be more easily understood in conjunction with the accompanying drawings. Figure 1 The genotype sequencing peak graphs of the SNP markers TT, TC, and CC of the present invention are displayed, and the blue background area is the SNP site. Specific embodiments

[0010] The embodiments of the present invention are described in detail below, and the present invention is further described in detail in conjunction with the embodiments. The embodiments herein are only used to illustrate the present invention and are not to be construed as limiting the present invention.

[0011] 1. Experimental samples The fasting body weights of 195 adult Macheng black goats from Hubei Jinyang (Macheng) Animal Husbandry Co., Ltd. were measured under the same feeding and management conditions and environmental conditions.

[0012] 2. Genomic DNA Extraction The jugular vein blood (5 mL / goat) of the above-mentioned experimental sample was collected using a disposable vacuum negative pressure blood collection tube (EDTA-K2 anticoagulation), and the genomic DNA of the blood samples of Macheng black goats was extracted according to the instructions of the blood genomic DNA extraction kit of Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0013] 3. Primer Design According to the sequence of goat FAM184B gene (Ensembl database gene sequence number: ENSCHIG00000018090), a pair of specific primers SEQ ID NO: 2 and SEQ ID NO: 3 were designed using Primer 6.0. The primers were synthesized by Beijing Qingke Biotechnology Co., Ltd. and were used to amplify a sequence where the second exon of the FAM184B gene is located. The amplified product was 616 bp, and the nucleotide sequence is shown in SEQ ID NO: 1 in the sequence table.

[0014] 4. Polymerase chain reaction (PCR) amplification and sequencing of the target sequence of the FAM184B gene of Macheng black goat (1) PCR amplification system (20 μL): 1 μL DNA (50 mg / mL), 1 μL primers SEQ ID NO: 2 and SEQ ID NO: 3 (100 uM), 10 μL 2×M5 HiPerplus Taq HiFi PCR mix (with blue dye) (purchased from Beijing Polymer Biotechnology Co., Ltd.), 7 μL ddH2O. The amplification program was as follows: 95°C pre-denaturation for 3 min, 94°C denaturation for 25 s, 55°C annealing for 25 s, 72°C extension for 9 s, 35 cycles, and 72°C final extension for 5 min. (2) The PCR amplification product was sent to Wuhan Jinkairui Biotechnology Co., Ltd. for sequencing. The sequencing results were analyzed using SnapGene software to determine the genotype of the individual at the 426 bp position of the nucleotide sequence shown in SEQ ID NO: 1 in the sequence list, such as Figure 1 As shown, the genotype of single peak T is TT, the genotype of single peak C is CC, and the genotype of double peak is TC. The base mutates from T to C, changing the encoded phenylalanine (Phe) into leucine (Leu).

[0015] 5. Association analysis between SNP markers of FAM184B gene and body weight in Macheng black goats One-way ANOVA in SPSS software was used to analyze the association between genotype and body weight. The specific linear analysis model is as follows: Y ij =μ+G i +E ij Where: Y ij is the individual phenotypic record; μ is the population mean; Gi is the genotype effect of each site; E ij is a random error.

[0016] 6. Analysis of the significance of weight differences among different genotypes of Macheng black goats The results of the analysis of the weight of different genotypes of Macheng black goats are shown in Table 1. As can be seen from Table 1, there are three genotypes at this locus. The difference in average weight between different genotypes was compared by one-way analysis of variance. It was found that the weight of Macheng black goats with TT and TC genotypes was significantly higher than that with CC genotype (p < 0.05). This shows that the TT and TC genotypes of this SNP locus can be used as an important criterion for judging the high weight of Macheng black goats. In the breeding of Macheng black goats, individuals with the genotype of TT at this locus can be retained and individuals with the genotype of CC at this locus can be eliminated. TC and TT type individuals can also be mated to obtain more TT genotype offspring individuals, thereby gradually improving the weight of the goat population. Table 1 Correlation between different genotypes of FAM184B gene mutation sites and body weight Note: Different letters in the same column indicate significant differences (p<0.05).

Claims

1. A molecular marker associated with the body weight of Macheng black goats, characterized in that: The molecular marker is located in the exon region of the FAM184B gene, and the specific nucleotide sequence is SEQ ID NO: 1, wherein the 426th base of the sequence has a T / C polymorphism.

2. A method for selecting goats according to the molecular marker of claim 1, wherein the weight of goat individuals with TT and TC genotypes at the molecular marker site is significantly higher than that of individuals with CC genotypes; The method comprises the following steps: (1) Extracting goat genomic DNA; (2) using two specific primers to perform PCR amplification to obtain a 616 bp amplification product, wherein the sequences of the two specific primers are SEQ ID NO: 2 and SEQ ID NO: 3; (3) sequencing the PCR amplification product to obtain a sequencing result; (4) determining the genotype of the goat individual to be tested at the molecular marker according to the sequencing result; (5) selecting goat individuals with the TT genotype of the above molecular marker for seed preservation; (6) The goat is a Macheng black goat.

3. Application of the molecular marker according to claim 1 in the breeding of Macheng black goat population, and selecting Macheng black goat individuals with TT genotype of the molecular marker for seed preservation.

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