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CN-121975945-A - SNP molecular marker related to sheep fertility and application thereof

CN121975945ACN 121975945 ACN121975945 ACN 121975945ACN-121975945-A

Abstract

The invention discloses an SNP molecular marker related to sheep fertility and application thereof. The SNP molecular marker comprises SNP1 and/or SNP2, wherein the SNP1 is positioned at 115747587-bit base of ARS-UI_ Ramb _V2.0 version 6 chromosome of sheep reference genome, the polymorphism is T/G, and the SNP2 is positioned at 41419566-bit base of ARS-UI_ Ramb _V2.0 version 22 chromosome of sheep reference genome, the polymorphism is A/G. The scheme of the invention provides the SNP molecular marker related to sheep fecundity, which can be used for efficiently and accurately identifying and screening sheep with high fecundity characters, has high mark parting quality, single copy and high polymorphism, and can be used for auxiliary breeding of sheep fecundity molecular markers.

Inventors

  • JIANG YOURU
  • JI ZHIBIN
  • HE LAN
  • YIN LIXIN
  • CAO MINGYUE
  • YE CHANGRONG
  • LI LE
  • TANG SHUNXUE

Assignees

  • 华智生物技术有限公司
  • 山东农业大学

Dates

Publication Date
20260505
Application Date
20260202

Claims (10)

  1. 1. A SNP molecular marker associated with sheep fertility, characterized in that said SNP molecular marker comprises SNP1 and/or SNP2; SNP1 is positioned at 115747587 base of the chromosome 6 of the sheep reference genome ARS-UI_ Ramb _V2.0 version, and the polymorphism is T/G; the SNP2 is positioned at 41419566 base of the 22 nd chromosome of the ARS-UI_ Ramb _V2.0 version of sheep reference genome, and the polymorphism is A/G.
  2. 2. A primer set for amplifying the SNP molecular marker according to claim 1.
  3. 3. The primer set of claim 2, wherein the primer set comprises: (1) Primer group for amplifying SNP1, comprising specific primers with sequences shown as SEQ ID NO.1 and SEQ ID NO.2, and/or, (2) A primer group for amplifying SNP2 comprises specific primers with sequences shown as SEQ ID NO.3 and SEQ ID NO. 4.
  4. 4. The primer set according to claim 3, wherein the specific primers are each linked to a different fluorescent group; Preferably, the fluorescent group is selected from FAM, JOE, VIC, HEX, ROX, CY or CY5.
  5. 5. A kit comprising the primer set of any one of claims 2 to 4.
  6. 6. Use of the SNP molecular marker of claim 1, the primer set of any one of claims 2-4 or the kit of claim 5 in any one of the following: 1) Detecting or aiding in the detection of sheep fertility; 2) Preparing a product for detecting or assisting in detecting sheep fertility; 3) Breeding sheep with high fertility character; 4) Preparing and breeding sheep products with high fertility characteristics; 5) Breeding sheep; 6) A product for sheep breeding was prepared.
  7. 7. A method for identifying or aiding in the identification of sheep fertility using the molecular marker of claim 1, comprising the steps of: s1, extracting genome DNA of sheep to be detected; s2, carrying out polymorphism detection on the SNP molecular markers on the genomic DNA extracted in the step S1, and judging the fertility of the sheep to be detected according to genotypes.
  8. 8. The method according to claim 7, wherein when the SNP molecular marker is SNP1, the sheep to be tested has a low fertility trait if the genotype detected by SNP1 is GG, and a high fertility trait if the genotype detected by SNP1 is TG or TT; When the SNP molecular marker is SNP2, the sheep to be detected has low fertility character if the genotype detected by the SNP2 is AA, and has high fertility character if the genotype detected by the SNP2 is AG or GG.
  9. 9. The method according to claim 7, wherein in step S2, SNP molecular markers are detected using KASP techniques; preferably, the composition of the KASP reaction mixture for detecting SNP molecular markers by using the KASP technique is as follows: Preferably, the amplification procedure for detecting SNP molecular markers by KASP technology is 92-95 ℃ for 13-17min, 92-95 ℃ for 18-22s,65 ℃ for 55-65s,8-12 cycles, 92-95 ℃ for 18-22s,55-58 ℃ for 55-65s,30-35 cycles.
  10. 10. A method for breeding sheep, comprising the step of selecting sheep having a high fertility trait for subsequent breeding by the method of any one of claims 7 to 9.

Description

SNP molecular marker related to sheep fertility and application thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to an SNP molecular marker related to sheep fertility and application thereof. Background Sheep (Ovis aries) are important economic animals worldwide, and the reproductive performance of sheep (Ovis aries) is directly related to the production efficiency and economic benefit of animal husbandry. Increasing lambing numbers, shortening lambing intervals, and improving breeding seasonality are the core goals of genetic improvement in sheep. With the rapid development of high-throughput sequencing technology and bioinformatics analysis methods, the genetic basis for controlling sheep reproductive traits is becoming increasingly well understood, and the identification of genes, quantitative Trait Loci (QTLs) and other genetic variations associated with reproductive and the accurate positioning of their positions on a reference genome is of great importance for developing modern breeding strategies such as molecular Marker Assisted Selection (MAS) and whole Genome Selection (GS). In recent years, the sheep industry gradually develops to a large-scale and intensive direction, but the sheep has obvious defects in the aspects of cultivation standardization, industrialization, economic benefit and the like. Particularly, in the current background of rapid development of barn feeding cultivation, the market demand for high-fertility sheep varieties is increasingly urgent, and improvement of industrial benefit through genetic improvement is urgently needed. Sheep fertility is mainly affected by two factors, namely the number of lambs born and the estrus cycle. Through long-term natural selection and artificial breeding, rich and various local sheep variety resources are formed. The fine breeds such as small-tailed han sheep and Hu sheep show the characteristics of perennial oestrus and high fertility, the lambing rate can reach more than 200%, while the breeds such as Tibetan sheep, xinjiang local sheep, yunnan local sheep and Mongolia sheep have the advantages of strong stress resistance, good adaptability and the like, but the problems of low fertility such as seasonal oestrus and high single embryo rate generally exist, and the improvement of the breeding benefit is severely restricted. The breeding method mainly comprises the steps of (1) interference by environmental factors (such as nutrition and management) to influence the accuracy of phenotype data, (2) large amount of manpower and material resources are needed to be input, the breeding cost is high, and (3) the breeding period is long (1-2 years), and the breeding efficiency is low. Genetic studies at home and abroad have reported some genes related to sheep reproduction, among which BMP15, GDF9 and BMPR1B are members of TGF- β superfamily, which play an indispensable role in follicular development and ovulation process, and mutation thereof is a major cause of multiple-embryo traits in many high-reproduction sheep varieties (e.g., small-tailed han sheep, booroola merino). The PRLR gene is associated with the prolactin signaling pathway and is thought to play a role in regulating the seasonal reproductive rhythm in sheep. Other genes such as TMEM154, CCNB2, etc., while their specific functional mechanisms are still under investigation, have been shown by a number of studies to be significantly associated with lambing or ovarian development. Some reproductive traits of sheep (e.g., total lambing number, conception rate) are complex quantitative traits commonly controlled by multiple genes. Quantitative Trait Locus (QTL) studies revealed that there are widely regions in sheep genomes that affect reproductive traits. These QTLs are distributed across multiple chromosomes, including 1,3, 5, 6, 10, 11, 15, 18, 20, etc. However, QTL localization intervals are often large, including a large number of genes, and it is difficult to directly target genes, but QTL localization is still an indispensable tool for mining trait-related genes. Whole genome association analysis (GWAS) utilizes high-density SNP markers that cover the whole genome, enabling more accurate identification of genetic loci associated with a trait of interest at the population level, typically at a higher resolution than traditional linkage analysis. GWAS analysis advances QTL localization from a broad chromosomal region to specific SNP sites and adjacent candidate genes. Not only can the effect of the known major genes in different groups be verified, but also new candidate genes with relatively small effects, such as FBXW8, NOS1 and the like, are continuously discovered. Therefore, the research on molecular genetic mechanism of sheep reproductive traits is of great practical significance. The main genes and key mutation sites for regulating and controlling the lambing number and the estrus cycle are mined, so that a target point ca