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CN-122012749-A - Application of FABP3 gene molecular marker g.2596A > C in Hu sheep molecular marker assisted breeding

CN122012749ACN 122012749 ACN122012749 ACN 122012749ACN-122012749-A

Abstract

The invention belongs to the technical field of molecular marker assisted breeding of Hu sheep, and particularly relates to application of FABP3 gene molecular marker g.2596A > C in molecular marker assisted breeding of Hu sheep. The invention takes Hu sheep as a research object, adopts a method of PCR amplification, direct sequencing of products and sequence analysis to analyze FABP3 gene polymorphism, and comprehensively analyzes the correlation between different genotypes and different growth traits of polymorphic sites. The analysis result shows that g.2596A > C is obviously related to the birth weight of Hu sheep, and the birth weight of AA genotype (P > 0.05) is obviously higher than that of CC genotype.

Inventors

  • HUANG XIN
  • GAO NING
  • GUO LIANGYONG
  • Shan Huili
  • JIANG JUNFANG
  • ZHENG KAIZHI
  • HE SANGANG

Assignees

  • 浙江省农业科学院
  • 湖州市农业科学研究院(湖州市农业科技发展中心)

Dates

Publication Date
20260512
Application Date
20260413

Claims (10)

  1. 1. A reagent for detecting and screening molecular markers of the birth weight of Hu sheep is characterized in that the molecular marker locus is g.2596A > C, wherein SNP loci are referenced to FABP3 gene positions on sheep reference genome GeneID= 100913166 and NC_056055.1:236035335-236042973, namely, 293A > C of a genome shown in SEQ ID NO. 1; in the molecular marker g.2596A > C, the birth weight of the AA genotype is significantly higher than that of the CC genotype.
  2. 2. A reagent for detecting molecular markers for screening of the birth weight of a Hu sheep according to claim 1, wherein the reagent comprises a primer pair for identifying the molecular markers.
  3. 3. The reagent for detecting molecular markers for screening of the birth weight of Hu sheep according to claim 1, wherein the nucleotide sequences of the primer pairs are as follows: F:5'- GCATCTTCTTCATTAGCAGGC-3'; R:5'- AGTGTTGGTCTCTGTTCGG-3'。
  4. 4. a kit comprising the reagent of any one of claims 1-3.
  5. 5. The reagent of any one of claims 1-3 or the kit of claim 4 for screening the hu sheep for birth weight auxiliary breeding, judging the hu sheep to be detected as a birth weight dominant candidate individual when the genotype is AA genotype, and judging the hu sheep to be detected as a non-birth weight dominant candidate individual when the genotype is AC genotype or CC genotype.
  6. 6. The application of an amplification product in screening of hu sheep birth weight assisted breeding is that the amplification product is obtained by amplification of the reagent according to any one of claims 1-3 or the kit according to claim 4, the nucleotide sequence of the amplification product is shown as SEQ ID NO.1, wherein the 293 th site is A or C, the hu sheep to be detected is judged to be a candidate individual of birth weight advantages when the genotype is AA genotype, and the hu sheep to be detected is judged to be a candidate individual of non-birth weight advantages when the genotype is AC genotype or CC genotype.
  7. 7. A method for screening the birth weight of a Hu sheep is characterized by comprising the following steps of extracting genomic DNA of the Hu sheep, carrying out PCR amplification by using a primer pair, detecting a molecular marker in an amplification product, and screening the birth weight of the Hu sheep, wherein the molecular marker locus is g.2596A > C in a FABP3 gene, namely, the 293A > C of a genome shown as SEQ ID NO. 1, when the genotype is an AA genotype, judging that the Hu sheep to be tested is a birth weight dominant candidate, and when the genotype is an AC genotype or a CC genotype, judging that the Hu sheep to be tested is a non-birth weight dominant candidate.
  8. 8. The method for screening the birth weight of Hu sheep according to claim 7, wherein the nucleotide sequences of the primer pairs are as follows: F:5'- GCATCTTCTTCATTAGCAGGC-3'; R:5'- AGTGTTGGTCTCTGTTCGG-3'。
  9. 9. The method for screening the birth weight of Hu sheep according to claim 7, wherein the PCR reaction system is 25.0. Mu.L, and comprises KOD OneTM PCR MASTER Mix-Blue 12.5. Mu.L, upstream and downstream primers each 0.2. Mu.L, template 1. Mu.L, ddH 2 O11.1. Mu.L.
  10. 10. The method for screening the birth weight of Hu sheep according to claim 7, wherein the PCR reaction is carried out by denaturing at 98 ℃ for 10 s, annealing at 61 ℃ for 30 s, and extending at 68 ℃ for 30 s, reacting for 34 cycles, and preserving at 4 ℃ after the end of PCR.

Description

Application of FABP3 gene molecular marker g.2596A > C in Hu sheep molecular marker assisted breeding Technical Field The invention belongs to the technical field of molecular marker assisted breeding of Hu sheep, and particularly relates to application of FABP3 gene molecular marker g.2596A > C in molecular marker assisted breeding of Hu sheep. Background The Hu sheep is an important local sheep variety in China, has the characteristics of high fertility, strong adaptability, faster early growth and the like, and has higher application value in mutton sheep production and variety improvement. Birth weight is one of important indexes for evaluating the early growth and development level of the lambs, is closely related to the body condition and the survival ability of the lambs in the primary stage, and can influence the subsequent growth performance and the group breeding effect. The traditional breeding mode is mainly selected and reserved according to phenotype record and pedigree information, judgment can be usually carried out after the target character is fully expressed, and the defects of long selection period, easiness in influence of breeding environment and management condition, low early identification efficiency and the like exist. Therefore, the molecular markers which are closely related to the birth weight of the Hu sheep and can be stably detected are mined and used for early auxiliary selection, and the molecular markers have important significance for improving the breeding efficiency of the Hu sheep, shortening the breeding period and improving the genetic improvement level of the population. Chinese patent No. CN111485026B discloses SNP locus, application, molecular marker and primer related to sheep birth weight, and discloses G/A polymorphic locus at 81799821 th chromosome of sheep to be related to sheep birth weight and can be used for auxiliary selection of early molecular marker of sheep birth weight. In addition, chinese patent No. CN114381531B discloses SNPs molecular marker g.43756G > A and application thereof in molecular marker assisted breeding of Hu sheep, and the patent uses a site related to Hu sheep body length characters for constructing a primer pair, a kit and a screening method, which shows that molecular marker assisted breeding of Hu sheep growth related characters by adopting specific SNP sites has a certain technical basis. In view of the fact that birth weight belongs to quantitative traits commonly affected by polygenes and genetic background, it is still necessary to further screen new candidate sites, and establish molecular markers with stronger pertinence and more convenient detection and application schemes thereof so as to meet the actual demands of molecular marker-assisted breeding of Hu sheep. FABP3 (fatty acid Binding Protein type 3, FATTY ACID Binding Protein 3), also known as cardiac fatty acid Binding Protein (H-FABP), is a fatty acid Binding Protein family member, expressed primarily in cardiac and skeletal muscle, and less in brown adipose tissue, nervous system and placenta. Plays a key role in the transport and metabolism of long chain fatty acids in cells, and provides a large amount of energy for biological activities. Studies have shown that knockout of the FABP3 gene in mice results in a disturbance of the plasma long chain fatty acid metabolism. However, research between the FABP3 gene and the Hu sheep body length character is not yet carried out, and the application of the FABP3 gene in the aspects of early screening of the Hu sheep growth character and molecular marker assisted breeding can be further explored. Disclosure of Invention In order to study the correlation of FABP3 gene polymorphism and Hu sheep growth character, genetic markers correlated with the growth character are obtained. The invention takes Hu sheep as a research object, adopts a method of PCR amplification, direct sequencing of products and sequence analysis to analyze FABP3 gene polymorphism, and comprehensively analyzes the correlation between different genotypes and different growth traits of polymorphic sites. The invention aims to provide a reagent for detecting molecular markers for screening Hu sheep body length, wherein the molecular marker locus is g.2596A > C, wherein SNP loci are respectively referenced to FABP3 gene positions on sheep reference genome GeneID= 100913166 and NC_056055.1:236035335-236042973, namely, the gene loci of the gene loci shown in SEQ ID NO. 1 are 293A > C; in the molecular marker g.2596A > C, the birth weight of the AA genotype is significantly higher than that of the CC genotype. Preferably, the reagent comprises a primer pair for identifying the molecular marker. Preferably, the nucleotide sequences of the primer pairs are as follows: F(SEQ ID NO:2):5'- GCATCTTCTTCATTAGCAGGC-3'; R(SEQ ID NO:3):5'- AGTGTTGGTCTCTGTTCGG-3'。 further, it is an object of the present invention to provide a kit comprising said reagent. Further, an object o