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CN-122012753-A - Promoter region SNP marker combination related to chicken egg laying performance, detection reagent and application thereof

CN122012753ACN 122012753 ACN122012753 ACN 122012753ACN-122012753-A

Abstract

The invention discloses a promoter region SNP marker combination related to chicken egg laying performance, a detection reagent and application thereof, and belongs to the technical field of molecular genetics. According to the invention, 3 SNP loci which are obviously related to egg laying performance are screened from a promoter of a CHST8 gene for the first time, are all positioned in a key regulation and control region of the promoter, have a functional basis for potentially influencing transcription factor combination and gene transcription activity, and all the 3 SNP loci show consistent effect trend among different genotypes, namely excellent genotypes show better key properties such as egg laying number. In molecular marker assisted breeding, the uniformity and stability of the effect direction are often of more practical significance, so that the promoter region SNP marker combination has important significance for breeding of high-yield laying hens.

Inventors

  • JIANG YUNLIANG
  • Shu Xinmei
  • ZHANG HONGRUI
  • KANG LI
  • SUN YI
  • WEI QINGQING

Assignees

  • 山东农业大学

Dates

Publication Date
20260512
Application Date
20260415

Claims (10)

  1. 1. A promoter region SNP marker combination related to chicken egg laying performance is characterized by comprising a first SNP marker and a second SNP marker; The nucleotide sequence of the first SNP marker is shown as SEQ ID NO.1, the 128 th base from the 5' end of the sequence is a SNP1 locus, and the base polymorphism is C or T; The nucleotide sequence of the second SNP marker is shown as SEQ ID NO.2, the 214 th base from the 5 'end of the sequence is a SNP2 site, the base polymorphism is T or C, the 276 th base from the 5' end of the sequence is a SNP3 site, and the base polymorphism is G or A.
  2. 2. The combination of SNP markers in promoter regions related to chicken egg laying performance according to claim 1, wherein the SNP1 locus is TT genotype, the SNP2 locus is CC genotype, and the SNP3 locus is AA genotype, which corresponds to quantitative multiple traits of egg laying.
  3. 3. A detection reagent is characterized by comprising a primer pair A for detecting a first SNP marker and a primer pair B for detecting a second SNP marker; The primer pair A consists of an upstream primer shown in SEQ ID NO.3 and a downstream primer shown in SEQ ID NO.4, and the primer pair B consists of an upstream primer shown in SEQ ID NO.5 and a downstream primer shown in SEQ ID NO. 6.
  4. 4. The detection reagent according to claim 3, wherein the detection reagent is a PCR sequencing kit or a KASP typing kit.
  5. 5. Use of the promoter region SNP marker combination related to chicken egg laying performance according to claim 1 or 2 for breeding hens with egg laying number multiple traits.
  6. 6. Use of the detection reagent according to claim 3 in (1) or (2) as follows: (1) Identifying egg laying characteristics of hens; (2) And breeding hens with the characteristics of multiple egg laying numbers.
  7. 7. The use of claim 6, wherein the egg laying trait comprises the number of eggs laid during the ascending period, the number of eggs laid during the peak period, or the total number of eggs laid at 43 weeks of age.
  8. 8. The use according to claim 6, wherein the method for breeding hens with a multi-character in egg production number is: The method comprises the steps of taking genome DNA of a hen to be detected as a template, carrying out PCR amplification on A by using a primer pair shown as SEQ ID NO.3 and SEQ ID NO.4 to obtain an amplification product A, carrying out PCR amplification on B by using a primer pair shown as SEQ ID NO.5 and SEQ ID NO.6 to obtain an amplification product B, sequencing the amplification product A and the amplification product B, identifying egg laying characteristics of the hen according to sequencing results, and breeding the hen with multiple egg laying quantity characteristics.
  9. 9. The use according to claim 8, wherein the number of eggs laid is determined to be multiple if the sequencing result of the amplification product A corresponds to the 128 th base of the sequence shown in SEQ ID NO.1 being of the TT genotype, the sequencing result of the amplification product B corresponds to the 214 th base of the sequence shown in SEQ ID NO.2 being of the CC genotype and the 276 th base being of the AA genotype.
  10. 10. The method according to claim 8, wherein the PCR amplification is performed using 2. Mu.L of genomic DNA, 25. Mu.L of 2X Phanta UniFi Master Mix, 2. Mu.L of each of the upstream and downstream primers, and up to 50. Mu.L of ddH 2 O; the PCR amplification procedure was 98℃pre-denaturation for 30sec, 98℃denaturation for 10sec,50℃ -53℃annealing for 10sec,72℃extension for 30sec, 35 cycles, and complete extension at 72℃for 5min after the end of the cycle.

Description

Promoter region SNP marker combination related to chicken egg laying performance, detection reagent and application thereof Technical Field The invention relates to the technical field of molecular genetics, in particular to a promoter region SNP marker combination related to chicken egg laying performance, a detection reagent and application thereof. Background The egg laying performance is a core economic character in chicken breeding, and mainly comprises indexes such as the age of open laying days, the number of eggs laid, the longest continuous laying days and the like, the genetic power of the egg laying performance is low, the egg laying performance is regulated and controlled by multiple genes and the environment together, and the traditional phenotype breeding has the limitations of long period, low efficiency, easiness in environmental interference and the like. As a variation form of a single base in a genome, single Nucleotide Polymorphism (SNP) has the advantages of high distribution density, strong genetic stability, automatic parting and the like, becomes a core molecular marker for poultry molecular breeding, and provides a key technical support for accurate genetic improvement of egg laying performance. In recent years, through researches such as whole genome association analysis, candidate gene method and the like, a large number of SNP markers which are obviously related to chicken egg laying performance are identified, key pathway genes such as endocrine regulation and follicular development are involved, and the SNP markers are involved in genetic regulation of egg laying properties by means of regulating gene expression quantity, changing protein structure and the like and are gradually applied to auxiliary selection of molecular markers of laying hens. However, the current research still has the problems of low functional SNP duty ratio, undefined SNP action mechanism of non-coding region, obvious difference of SNP breeding values of different regions and the like, and restricts the accurate development of molecular breeding. The gene promoter region is a core regulatory region located upstream of the transcription initiation site, and directly controls the transcription initiation efficiency and expression level of the gene. Compared with the gene exon region, the intron region and the 3'UTR/5' UTR region molecular marker, the functional effect of the promoter region SNP is clear, the transcription factor binding affinity can be directly changed, the gene expression is regulated and controlled from the source, the problem of functional ambiguity caused by synonymous mutation and neutral variation is avoided, the marker specificity is high, the genetic stability is high, the coordinated regulation of the multi-egg-laying characteristics can be realized, and the marker has remarkable advantages in molecular breeding of egg-laying performance. The current research of SNP in promoter region aiming at chicken egg laying performance is still in a starting stage, the number of high-quality functional markers is limited, SNP in promoter region of key gene of egg laying is deeply excavated, the regulation and control mechanism is analyzed, and efficient breeding application is developed, so that the method has important theoretical and practical significance for improving the genetic improvement efficiency of laying hens and cultivating new varieties of high-quality laying hens. CHST8 (Carbohydrate sulfotransferase) belongs to one of the members of the carbohydrate-sulfate transferase family, and is primarily involved in the sulfation modification process of glycoproteins and glycolipids, which play an important role in extracellular matrix structure maintenance, intercellular signaling, and regulation of cellular function. At present, the research on the CHST8 gene in poultry is still limited, and no report on the correlation between SNP markers of a promoter region of the CHST8 gene and chicken egg laying performance is yet seen. Disclosure of Invention Aiming at the prior art, the invention aims to provide a promoter region SNP marker combination related to chicken egg laying performance, a detection reagent and application thereof. In order to achieve the above purpose, the invention adopts the following technical scheme: in a first aspect of the present invention, there is provided a promoter region SNP marker set related to chicken egg laying performance, comprising a first SNP marker and a second SNP marker; The nucleotide sequence of the first SNP marker is shown as SEQ ID NO.1, the 128 th base from the 5' end of the sequence is a SNP1 locus, and the base polymorphism is C or T; The nucleotide sequence of the second SNP marker is shown as SEQ ID NO.2, the 214 th base from the 5 'end of the sequence is a SNP2 site, the base polymorphism is T or C, the 276 th base from the 5' end of the sequence is a SNP3 site, and the base polymorphism is G or A. The nucleotide of the first SNP marker