SNP marker for improving body weight of goat and application thereof

Abstract

The application discloses a SNP marker related to goat weight and application thereof. The marker is located on an exon of a FAM184B gene on a goat chromosome 6, and a specific SNP marker is a T / C base mutation at the 426th bp in SEQ ID NO: 1 in the sequence listing; the weight of individuals with TT and TC genotypes is significantly higher than that of individuals with the CC genotype. The application takes Macheng black goats as research objects, amplifies the DNA sequence of the goat FAM184B gene by using PCR, analyzes the influence of mutation site polymorphism on the weight of the goat, and can be used for breeding the Macheng black goats, accelerates the breeding of the Macheng black goat breed taking the Macheng black goat as a breeding material, and provides a basic marker resource for marker-assisted selection breeding of the weight trait of the goat.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of animal molecular breeding, and relates to application of a SNP marker related to a goat weight trait in breeding of Macheng Black goats. BACKGROUND

[0002] The purpose of meat-producing livestock and poultry production is to produce meat, and the larger the individual weight, the more meat is produced, so weight is a key indicator for evaluating goat productivity and economic benefits. The phenotype of a trait is controlled by genes, so molecular marker-assisted selection according to genes controlling the trait has the advantages of simple operation and early selection, and can effectively shorten the breeding period and improve breeding efficiency.

[0003] A large number of omics analyses show that the Family with sequence similarity 184 member B (FAM184B) gene is related to livestock body type and weight traits. Based on GWAS analysis, it has been determined that the gene affects horse body type, donkey body height, 14-month-old body weight and muscle growth traits of sheep, and body weight of chickens and meat pigeons, and it is found that one SNP of the gene affects horse body length and hock circumference, and seven SNPs are significantly related to carcass weight. In summary, the FAM184B gene is closely related to livestock and poultry growth traits, and is related to muscle growth and bone development.

[0004] Macheng Black goats are excellent black goat meat breeds in the Dabie Mountains, and have growth characteristics such as fast growth rate and good fattening ability. Based on whole-genome selection signal analysis, Macheng Black goats were divided into high weight and low weight groups, and positive selection signals were detected, and it was found that one positive selection SNP marker of the FAM184B gene affected weight, providing a new marker resource for molecular marker-assisted selection breeding of Macheng Black goat weight traits. SUMMARY

[0005] The application aims to provide a technical method for goat weight selection.

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

[0007] The application provides a primer pair for detecting the SNP marker in claim 1. According to the embodiment of the application, the primer has the nucleotide sequence shown in SEQ ID NO: 2 and SEQ ID NO: 3, the primer pair of the application can effectively perform PCR amplification on the fragment where the above-mentioned SNP marker related to the body weight of the goat to be detected, and the detection of the SNP marker can be effectively realized through sequencing, the genotype of the SNP marker of the goat to be detected is determined, and the body weight of the goat to be detected can be effectively predicted. Specifically, the body weight of the individual with TT and TC genotypes of the SNP site is significantly higher than that of the individual with CC genotype, and can be used as an important standard for judging the body weight of the goat. In the breeding work of the goat, the individual with the genotype TT of the SNP site can be reserved for breeding, the individual with the genotype CC of the SNP marker site can be eliminated, and the individual with the genotype CT can be mated with the individual with the genotype TT to obtain more offspring with the genotype TT, so that the excellent breed of the goat is bred at low cost and high accuracy, and the body weight of the goat population is gradually increased.

[0008] The application has the following beneficial effects: (1) the SNP marker provided by the application is significantly related to the body weight of Macheng black goat, and the body weight of the Macheng black goat with TT and TC genotypes is significantly higher than that of the Macheng black goat with CC genotype; (2) the SNP marker can be used for assisted selection of the body weight of the Macheng black goat, and the Macheng black goat with high body weight is screened out, which has important practical application value for further increasing the body weight of the Macheng black goat and breeding the breed (or strain) using the Macheng black goat as a material. BRIEF DESCRIPTION OF DRAWINGS

[0009] The above aspects of the application will be easier to understand from the description of the embodiments in combination with the drawings, Figure 1 The SNP marker TT, TC and CC of the application is shown in the sequencing peak graph, and the blue background area is the SNP site. DETAILED DESCRIPTION

[0010] The embodiments of the application are described in detail below, and the application is further described in combination with the embodiments, and the embodiments herein are only used for describing the application and cannot be understood as limiting the application.

[0011] 1. Experimental samples

[0012] The fasting body weight of 195 adult Macheng black goats of Hubei Jinyang (Macheng) Animal Husbandry Co., Ltd. is determined, and the feeding management conditions and environmental conditions of the goats are the same.

[0013] 2. Genomic DNA extraction

[0014] The jugular vein blood (5 mL / one) of the above-mentioned 1 experimental sample was collected by using a disposable vacuum negative pressure blood collection tube (EDTA-K2 anticoagulant), and the genomic DNA of the Macheng black goat blood sample was extracted according to the blood genomic DNA extraction kit instruction book of Tian Gen Biochemical Technology (Beijing) Co., Ltd.

[0015] 3. Primer design

[0016] According to the sequence of the 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 by using Primer 6.0, and the primers were synthesized by Beijing Qikang Biological Technology Co., Ltd. and used to amplify a sequence in which the 2nd exon of the FAM184B gene was located. The amplification product was 616 bp, and the nucleotide sequence was shown in SEQ ID NO: 1 in the sequence listing.

[0017] 4. Polymerase chain reaction (PCR) amplification of Macheng black goat FAM184B gene target sequence and sequencing

[0018] (1) PCR amplification system (20 μL): 1 μL of DNA (50 mg / mL), 1 μL of primers SEQ ID NO: 2 and SEQ ID NO: 3 (100 uM) each, 10 μL of 2xM5 HiPerplus Taq HiFi PCR mix (with blue dye) (purchased from Beijing Junxing Biological Technology Co., Ltd.), and 7 μL of ddH2O. The amplification program was as follows: 95℃ pre-denaturation for 3 min, 94℃ denaturation for 25 s, 55℃ annealing for 25 s, 72℃ extension for 9 s, 35 cycles, and 72℃ final extension for 5 min.

[0019] (2) The PCR amplification product was sent to Wuhan Jin Kai Rui Biological Technology Co., Ltd. for sequencing. The sequencing results were analyzed by using SnapGene software, and the genotype of the individual at the 426 bp site of the nucleotide sequence shown in the sequence listing SEQ ID NO: 1 was determined. As shown in Figure 1 , the genotype of single-peak T was TT, the genotype of single-peak C was CC, and the genotype of double-peak was TC. The base was mutated from T to C, which changed the encoded phenylalanine (Phe) to leucine (Leu).

[0020] 5. Association analysis of Macheng black goat FAM184B gene SNP marker and body weight

[0021] The association analysis between genotypes and body weight was performed by using single factor variance analysis in SPSS software. The specific linear analysis model is as follows:

[0022] Y ij = μ + Gi +E ij

[0023] wherein: Y ij is the individual phenotype record; μ is the population mean; G i is the genotype effect of each locus; E ij is the random error.

[0024] 6. Analysis of the significance of the weight difference between different genotypes of Macheng black goats

[0025] 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 the locus. Single factor variance analysis was used to compare the weight difference between different genotypes, and it was found that the weight of Macheng black goats with TT and TC genotypes was significantly higher than that of goats with CC genotype (p<0.05). This indicates that TT and TC genotypes of the SNP locus can be used as an important standard for judging high weight of Macheng black goats. In the breeding work of Macheng black goats, individuals with genotype TT at the locus can be retained, and individuals with genotype CC at the locus can be eliminated. Alternatively, TC and TT individuals can be mated to obtain more offspring with genotype TT, so as to gradually increase the weight of the goat population.

[0026] Table 1 Correlation between different genotypes of FAM184B gene mutation locus and weight

[0027]

[0028] Note: Different superscripts in the same column indicate significant difference (p<0.05).

Claims

1. A method for selecting high-weight goats using a molecular marker related to the weight of Macheng black goats, wherein 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, and the weight of goat individuals with the TT and TC genotypes at the molecular marker site is significantly higher than that of individuals with the CC genotype; The method comprises the following steps: (1) extracting goat genomic DNA; (2) obtaining a 616bp amplification product by PCR amplification using two specific primers, 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 to-be-tested goat individual at the molecular marker according to the sequencing result; (5) selecting a goat individual with a TT genotype of the molecular marker for breeding; (6) the goat is a Macheng Black goat.

2. Application of the molecular marker in claim 1 in the breeding of a high-weight Macheng Black goat population, wherein a Macheng Black goat individual with a TT genotype of the molecular marker is selected for breeding.

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