SNP marker affecting yean trait of Huyang sheep (sheep raised in Zhejiang Province) and application of SNP marker

A trait and molecular marker technology, applied in the field of molecular biotechnology and molecular markers, can solve the problems of increasing the difficulty of SNP molecular markers, inaccurate positioning, large DNA requirements, etc., to increase core competitiveness, huge profit potential, and speed up breeding. effect of the process

Active Publication Date: 2018-08-17
SOUTH CHINA AGRI UNIV
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  • Abstract
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  • Claims
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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 QTLs c

Method used

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  • SNP marker affecting yean trait of Huyang sheep (sheep raised in Zhejiang Province) and application of SNP marker
  • SNP marker affecting yean trait of Huyang sheep (sheep raised in Zhejiang Province) and application of SNP marker
  • SNP marker affecting yean trait of Huyang sheep (sheep raised in Zhejiang Province) and application of SNP marker

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Experimental program
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Effect test

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 is raised in high-bed sheds throughout the year, and the rations are fed 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...

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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Abstract

The invention belongs to the fields of molecular biotechnology and molecular marker technology, and relates to applications of an SNP molecular marker in researches on yean trait of sheep and in sheepbreeding. The site of the sheep SNP molecular marker is shown as SEQ ID NO.1, and nucleic acid single-base mutation at the 193rd site of the sequence fragment is named as G765A. The SNP molecular marker corresponds to G>A mutation at the 765bp of a coding region sequence of a BMPR-IB gene (accession number: NC-019463.2) on the No.6 sheep chromosome, which is issued, on GeneBank. According to theinvention, by optimizing a dominant allele type of the SNP molecular marker, a genetic progress of yean trait of the Huyang sheep can be accelerated and breeding time of reproduction trait of the Huyang sheep can be shortened, so that reproduction cost is reduced, and economic benefits of mutton sheep breeding industry are 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] 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...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124
Inventor 李耀坤杨新月柳广斌刘德武孙宝丽郭勇庆邓铭
Owner SOUTH CHINA AGRI UNIV
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