SNP marker affecting litter size trait of hu-sheep and application thereof

A technology of traits and molecular markers, which is applied in the field of molecular biotechnology and molecular markers, can solve the problems of increasing the difficulty of SNP molecular markers, the large amount of DNA required, and the complicated detection technology, so as to increase core competitiveness, increase genetic progress, and mine breeding potential effect

Active Publication Date: 2018-06-01
SOUTH CHINA AGRI UNIV
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Problems solved by technology

Patent CN104099330A discloses that the allelic mutation occurs at position 713 of the LHβ gene fragment of sheep (breeds include Dorper and Hu sheep), which affects the number of lambs born in Mianyang
[0005] However, there are still some problems in molecular breeding at present, mainly including: the limitation of marker genetic map; the polymorphism frequency of molecular markers linked to breeding target traits is low; The degree of automation is low, and the required equipment and reagents are expensive
However, in gene mapping, due to the limitations of current mapping materials, there is often the problem of inaccurate positioning, and in the technology of QTL mapping of quantitative trait loci, not all target QTL...

Method used

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  • SNP marker affecting litter size trait of hu-sheep and application thereof
  • SNP marker affecting litter size trait of hu-sheep and application thereof
  • SNP marker affecting litter size trait of hu-sheep and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A total of 198 Hu sheep ewes with complete lambing records were selected, and 2 mL of blood was drawn from the jugular vein for anticoagulant treatment, 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 is raised in high-bed sheds throughout the year, and the rations are fed according to the uniform feeding standard. Eat and drink.

[0037] (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 sa...

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 791bp nucleotide sequence on chromosome 6, and the upstream and downstream primers for sequence amplification are based on reports on GenBank The sequence of the sheep BMPR-IB gene (accession number: NC-019463.2), using Primer Premier 5.0 software to design primers for exon 1 of the BMPR-IB gene, and amplify the partial sequence of the 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'-TTTTAGGGCTCCATGTACTCC-3';

[0052] SEQ ID NO.3

[0053] Downstream primer PCR-R: 5'-TTTCCTCTGGGTGCTGTG-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 polymera...

Embodiment 3

[0063] Example 3 SNP site T-87C effect analysis of molecular markers

[0064] The invention provides a SNP marker that can significantly increase the lambing size 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 CC-type individuals, and the average lamb size of each breeding ewe can be increased by 0.27, which shows that the breeding of breeding ewes with high reproductive performance , which is of great significance to reduce the cost of reproduction, improve the economic benefits of sheep farming 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 CC individuals and CT individuals (P<0.05), and by optimizing the dominant allele (C) ...

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Abstract

The invention belongs to the technical fields of molecular biology and molecular marker, and particularly relates to application of a sheep SNP molecular marker to sheep little size trait research andsheep breeding. The locus of the sheep SNP molecular marker is as shown in SEQ ID NO.1, and mutation of nucleic acid single base positioned in the 343th position of a sequence segment is named as T-87C. The SNP molecular marker corresponds to T-to-C mutation in the 87bp position of a coding sequence of BMPR-IB gene (accession number: NC_019463.2) on sheep No.6 chromosome released on GeneBank. Through optimization of superiority allelomorphic gene of the SNP molecular marker, genetic progress of litter size trait of hu-sheep can be improved, and the breeding time of hu-sheep with breeding trait is shortened, so that the reproductive cost is reduced, and the economic benefit of the mutton sheep breeding industry is effectively improved.

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] Lamb size is the most important reproductive trait of sheep. It is affected by factors such as genes and environment, and its economic benefits are very significant. It is the main factor affecting the development of mutton sheep industry. Therefore, lambing traits are the focus of research on genetic improvement of breeding sheep. It is difficult to quickly improve the reproductive performance of mutton sheep by conventional breeding methods, but marker assisted selection (MAS) breaks this technical barrier and can eliminate the selection of phenotypic traits by factors such as age, sex, and environment. interference, thus making up for the lack of conventional phenotype selection (Wang Baichua...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 刘德武杨新月李斌柳广斌李耀坤郭勇庆邓铭
Owner SOUTH CHINA AGRI UNIV
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