A snp marker affecting lambing traits of Hu sheep and its application

Abstract

The invention belongs to the technical fields of molecular biotechnology and molecular markers, and relates to the application of a sheep SNP molecular marker in the study of sheep lambing traits and sheep breeding. The goat SNP molecular marker site is shown in SEQ ID NO.1, and the nucleic acid single base mutation at position 193 of the sequence fragment is named: G765A. The SNP molecular marker corresponds to the G>A mutation at the 765th bp of the coding region sequence of the BMPR‑IB gene (accession number: NC_019463.2) on the sheep chromosome 6 that has been published on GeneBank. By optimizing the dominant allelic genotype of the SNP molecular marker, the present invention can accelerate the genetic progress of lambing traits of Hu sheep, shorten the breeding time of reproductive traits of Hu sheep, thereby reducing the cost of reproduction, and effectively improving the economic benefits of the mutton sheep breeding industry .

Description

technical field

[0001] The invention belongs to the field of molecular biotechnology and molecular marker technology, and in particular relates to the application of a Hu sheep SNP molecular marker in the study of sheep lambing traits and sheep breeding. Background technique

[0002] For breeding stock, fecundity is productivity, and fecundity is directly related to the economic effect of mutton sheep industry. Reproductive performance mainly includes the total number of lambs born, the number of live lambs born, birth weight, newborn litter weight, weaned litter weight and other indicators. Low lambing rate is the main bottleneck restricting the development of high-quality and efficient sheep raising industry. It is difficult to quickly improve the reproductive performance of mutton sheep by adopting conventional breeding methods. However, it is very important to study the breeding method of improving lambing rate from the genetic nature. Sheep scientific and technological...

Examples

Embodiment 1

[0035] In the embodiment, 811 Hu sheep ewes with complete lambing records were selected, and 2 mL of jugular vein blood was drawn and anticoagulated, and stored at -20°C for later use. The Hu sheep used in this research are all from the Liangdong sheep breeding farm of Wenshi Xinwang Sheep Industry Co., Ltd., Xinxing County, Yunfu City, Guangdong Province. The farm adopts high-bed house feeding throughout the year, and feeds rations according to the uniform feeding standard. Eat and drink.

[0036] (1) The extraction of Hu sheep blood genomic DNA refers to the blood genomic DNA extraction kit (purchased from Tiangen Biochemical Technology Co., Ltd.), and is detected by 1% agarose gel electrophoresis without dispersion or tailing phenomenon; A spectrophotometer is used to detect the quality and concentration of the extracted DNA. DNA samples with A260 / 280 ratio of 1.8 to 2.0 and A260 / 230 ratio of 1.7 to 1.9 were judged as qualified. Finally, the qualified DNA samples were uni...

Embodiment 2

[0049] (1) Amplification of the target fragment containing the SNP site significantly related to the lambing traits of Hu sheep: the target fragment is a 261bp nucleotide sequence on chromosome 6, and the upstream and downstream primers for sequence amplification are based on reports on GenBank Sheep BMPR-IB gene sequence (accession number: NC-019463.2), using Primer Premier 5.0 software to design primers for exon 8 of BMPR-IB gene, and amplify the partial sequence of sheep BMPR-IB gene. The primers were synthesized by Huada Biotechnology Co., Ltd., and the specific sequences are as follows:

[0050] SEQ ID NO.2

[0051] Upstream primer PCR-F: 5'-GTCTGCTGTATTGGCACACACAT-3';

[0052] SEQ ID NO.3

[0053] Downstream primer PCR-R: 5'-ACCATCTGAATTTGCTTTGC AT-3'.

[0054] (2) PCR amplification system and program setting

[0055] PCR reaction system: 10 μL system contains 1 μL of genomic DNA template, 0.2 μL of upstream and downstream primers, 5 μL of TaqDNA polymerase, supple...

Embodiment 3

[0063] Example 3 SNP site G765A effect analysis of molecular markers

[0064] The invention provides a SNP marker that can significantly increase the lambing number of Hu sheep, and the SNP is used for marker-assisted selection, which can greatly increase the breeding process of the Hu sheep's reproductive performance.

[0065] In the present invention, the molecular marker SNP sites that affect the lamb size of sheep are all selected into AA-type or GG-type individuals, and the average lamb size of each breeding ewe can be increased by 0.38 and 0.39 respectively, which shows that the reproductive performance is high The breeding of ewes is of great significance to reduce the cost of breeding, improve the economic benefits of sheep breeding and promote the rapid development of my country's mutton sheep industry. Among the individuals marked by this SNP, there is a significant difference in litter size between individuals of type AA and GG and individuals of type GA (P<0.05). T...

technical field

[0001] The invention belongs to the field of molecular biotechnology and molecular marker technology, and in particular relates to the application of a Hu sheep SNP molecular marker in the study of sheep lambing traits and sheep breeding. Background technique

[0002] For breeding stock, fecundity is productivity, and fecundity is directly related to the economic effect of mutton sheep industry. Reproductive performance mainly includes the total number of lambs born, the number of live lambs born, birth weight, newborn litter weight, weaned litter weight and other indicators. Low lambing rate is the main bottleneck restricting the development of high-quality and efficient sheep raising industry. It is difficult to quickly improve the reproductive performance of mutton sheep by adopting conventional breeding methods. However, it is very important to study the breeding method of improving lambing rate from the genetic nature. Sheep scientific and technological...

Examples

Embodiment 1

[0035] In the embodiment, 811 Hu sheep ewes with complete lambing records were selected, and 2 mL of jugular vein blood was drawn and anticoagulated, and stored at -20°C for later use. The Hu sheep used in this research are all from the Liangdong sheep breeding farm of Wenshi Xinwang Sheep Industry Co., Ltd., Xinxing County, Yunfu City, Guangdong Province. The farm adopts high-bed house feeding throughout the year, and feeds rations according to the uniform feeding standard. Eat and drink.

[0036] (1) The extraction of Hu sheep blood genomic DNA refers to the blood genomic DNA extraction kit (purchased from Tiangen Biochemical Technology Co., Ltd.), and is detected by 1% agarose gel electrophoresis without dispersion or tailing phenomenon; A spectrophotometer is used to detect the quality and concentration of the extracted DNA. DNA samples with A260 / 280 ratio of 1.8 to 2.0 and A260 / 230 ratio of 1.7 to 1.9 were judged as qualified. Finally, the qualified DNA samples were uni...

Embodiment 2

[0049] (1) Amplification of the target fragment containing the SNP site significantly related to the lambing traits of Hu sheep: the target fragment is a 261bp nucleotide sequence on chromosome 6, and the upstream and downstream primers for sequence amplification are based on reports on GenBank Sheep BMPR-IB gene sequence (accession number: NC-019463.2), using Primer Premier 5.0 software to design primers for exon 8 of BMPR-IB gene, and amplify the partial sequence of sheep BMPR-IB gene. The primers were synthesized by Huada Biotechnology Co., Ltd., and the specific sequences are as follows:

[0050] SEQ ID NO.2

[0051] Upstream primer PCR-F: 5'-GTCTGCTGTATTGGCACACACAT-3';

[0052] SEQ ID NO.3

[0053] Downstream primer PCR-R: 5'-ACCATCTGAATTTGCTTTGC AT-3'.

[0054] (2) PCR amplification system and program setting

[0055] PCR reaction system: 10 μL system contains 1 μL of genomic DNA template, 0.2 μL of upstream and downstream primers, 5 μL of TaqDNA polymerase, supple...

Embodiment 3

[0063] Example 3 SNP site G765A effect analysis of molecular markers

[0064] The invention provides a SNP marker that can significantly increase the lambing number of Hu sheep, and the SNP is used for marker-assisted selection, which can greatly increase the breeding process of the Hu sheep's reproductive performance.

[0065] In the present invention, the molecular marker SNP sites that affect the lamb size of sheep are all selected into AA-type or GG-type individuals, and the average lamb size of each breeding ewe can be increased by 0.38 and 0.39 respectively, which shows that the reproductive performance is high The breeding of ewes is of great significance to reduce the cost of breeding, improve the economic benefits of sheep breeding and promote the rapid development of my country's mutton sheep industry. Among the individuals marked by this SNP, there is a significant difference in litter size between individuals of type AA and GG and individuals of type GA (P<0.05). T...

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