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Molecular marker for sheep multifetal traits and its application

A technology of molecular markers and traits, applied in the field of molecular biology and molecular breeding, to speed up the breeding process, promote economic benefits, and promote the effect of improving

Active Publication Date: 2021-07-23
黑龙江省农业科学院畜牧兽医分院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research report on the existence of molecular markers of bone morphogenetic protein 2 (BMP2) related to the multiple births of Small-tailed Han sheep

Method used

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  • Molecular marker for sheep multifetal traits and its application
  • Molecular marker for sheep multifetal traits and its application
  • Molecular marker for sheep multifetal traits and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Discovery of polymorphic sites of BMP2 gene in Small-tailed Han sheep

[0024] The experimental subjects were selected from 300 healthy small-tailed Han sheep with complete lambing records in the applicant's own farm, and 2ml of blood was drawn from the jugular vein, treated with anticoagulation, and stored at -20°C until use.

[0025] (1) Extraction of genomic DNA. Genomic DNA was extracted using the Blood Genomic DNA Extraction Kit from Tiangen Biochemical Technology Co., Ltd., and no tailing or smearing was detected by 1% agarose gel electrophoresis. The DNA was detected with an ultraviolet spectrophotometer, and the A260 / 280 ratio was in the range of 1.8-2.0, and the A260 / 230 ratio was in the range of 1.7-1.9, and the quality of the sample was qualified. Dilute the sample to 50ng / μl and freeze it at -20°C.

[0026] (2) Exon sequencing of BMP2 gene. Randomly select 20 small-tailed Han sheep DNA samples to construct pool DNA for exome sequencing. The gen...

Embodiment 2

[0028] Example 2 Genotype frequency and allele frequency of polymorphic loci

[0029](1) PCR amplifies the DNA fragment of BMP2 exon 2 containing the mutation site. Primers were designed using Primer Premier 5.0 to amplify the DNA fragment containing the mutation site and the restriction endonuclease site analysis was performed. Upstream primer F: 5'-TTCTAGCGTTGCTGCTTCCC-3' (SEQ ID NO: 2); Downstream primer R: 5'-TTCGTGGTGAAAGCTGCG-3' (SEQ ID NO: 3). PCR reaction system: 10 μl system contains 1 μl of template, 0.2 μl of upstream and downstream primers, 5 μl of Taq DNA polymerase, and milli-Q water for the rest; PCR reaction program: pre-denaturation at 94°C for 2 minutes, 35 cycles (denaturation at 94°C for 30s , annealed at 60°C for 30s, extended at 71°C for 30s), extended at 71°C for 5min, and stored at 4°C until use. The PCR product is a 320bp DNA fragment (SEQ ID NO: 4), wherein the 47th position is a single nucleotide mutation site (C>T). After analysis, the mutation s...

Embodiment 3

[0036] Example 3 Correlation analysis between different genotypes and multiple birth traits

[0037] The GLM (General LinearModel) program in the SPSS 2.0 statistical analysis software was used to cooperate with the constructed single-site effect model to carry out statistical analysis on the molecular marker sites, and the LSD and Dunnett's T3 methods were used to perform multiple comparisons between the means to study the parity and genotype on the multifetal traits, the linear model used for statistical analysis is as follows:

[0038] the y jkl =μ+P j +G k +e jkl

[0039] where: y jkl is the recorded value of individual litter number; μ is the average value of the group; P j is the fixed effect of the jth parity; G k is the fixed effect of the kth genotype; e jkl is a random residual effect. All data are presented in the form of mean ± standard deviation (Mean ± SD). Table 2 shows the relationship between the genotype of the molecular marker C76T on exon 2 of BMP2...

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Abstract

The invention belongs to the fields of molecular biology and molecular breeding. Specifically, the present invention provides a SNP molecular marker on the BMP2 gene of Small-tailed Han sheep related to multiple fecundity traits, a detection method of the molecular marker and the application of the method in improving the reproductive performance of sheep and breeding multiple fecundity varieties. The molecular marker is the polymorphic site at position 76 of exon 2 of BMP2 gene, denoted as C76T. Statistical analysis showed that the molecular marker C76T was significantly correlated with multiple birth traits, and the allele T had a significant additive effect. Therefore, in the process of breeding and breeding sheep, the SNP molecular marker can be used to assist in the selection of CT-type or TT-type individuals with high fecundity, thereby accelerating the breeding process of sheep reproductive performance.

Description

technical field [0001] The invention belongs to the fields of molecular biology and molecular breeding. Specifically, the present invention provides a SNP molecular marker related to BMP2 gene of Small-tailed Han sheep and multiple fecundity traits, a detection method of the molecular marker and the application of the method in improving the reproductive performance of sheep and breeding multiple fecundity breeds. Background technique [0002] Reproductive performance is an important economic trait of sheep, especially in the process of shifting the breeding direction of sheep from the past single use of wool and wool to meat and wool or meat, the pursuit of higher lambing rate and more fetuses Lamb size is of great economic importance. The reproductive performance of sheep is mainly affected by three factors: multiple birth rate, conception rate and survival rate, among which multiple birth rate has the largest variable. Sheep are single-born mammals, but some sheep breed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 刘学峰张立春李信涛海龙李旭业郭立宏姜明生王嘉厚吴宪郝彩虹杜军吕雄杰孙继权穆文利王锋尤海洋李莉郭文凯杨淑萍刘丽秋刘秀玲王佳王宇董扬张超
Owner 黑龙江省农业科学院畜牧兽医分院
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