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Genetic marker related to porcine semen quality traits and application

A quality trait and genetic marker technology, applied in the field of pig genetic marker screening, can solve the problems of slow progress and low boar heritability

Inactive Publication Date: 2016-04-27
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Routine selection has been slow due to low heritability of boar reproductive traits

Method used

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  • Genetic marker related to porcine semen quality traits and application
  • Genetic marker related to porcine semen quality traits and application
  • Genetic marker related to porcine semen quality traits and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: The acquisition of pig RNF4 gene fragments and the establishment of polymorphism detection methods

[0026] Log in to the NCBI database, download the porcine RNF4 gene sequence as the target sequence, and use the PrimerPremier5 software to design primers. The primer sequence is shown in SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence table, and the specific sequence is as follows: forward primer RNF4-F: AGAACGCTAACACTTGCC, reverse primer RNF4-R: CACGGTGACCTCTGACTC. The amplified region includes the 649bp genomic nucleotide sequence of the 3' untranslated region of the pig RNF4 gene, such as figure 1shown. The PCR reaction system was 25 μl, in which the template DNA was 50 ng, the concentration of dNTPs was 200 μmol / L, the concentration of each primer was 0.4 μmol / L, 3 U of TaqDNA polymerase (Biostar International, Canada), and deionized water was added to a total volume of 25 μl; Reaction program: pre-denaturation at 94°C for 4 minutes; then denaturation at ...

Embodiment 2

[0028] Example 2: Detection of polymorphism distribution of genetic markers screened by the present invention in different pig herds

[0029] The extraction of pig genome DNA (sample is shown in Table 1) is carried out with reference to the method reported in 1999 in Xiong Yuan's "Introduction to Pig Biochemistry and Molecular Genetic Experiments" China Agricultural Press.

[0030] The PCR-AluI-RFLP polymorphism of the porcine RNF4 gene was detected in three foreign blood-related pig populations, and the detection results are shown in Table 1. The results showed that there were three genotypes in Duroc, Large White and Landrace pigs; in the Duroc population, the C allele frequency was 0.84, which was the dominant allele; in the Large White pig population, the T allele The frequency is 0.78, which is the dominant allele; in the Landrace pig population, the C allele frequency and T allele frequency are 0.44 and 0.56, respectively, which are alleles with no obvious advantages (se...

Embodiment 3

[0034] Example 3: Application of the association analysis between the genetic markers screened by the present invention and pig semen quality traits

[0035] In order to determine whether the pig RNF4 gene PCR-AluI-RFLP is related to the difference in the quality of pig semen, the Duroc pigs, Large White pigs and Landrace pigs bred in Guangxi Yangxiang Pig Farm were selected as test materials, and the AluI-RFLP established in Example 1 was used. Methods The polymorphism was detected, and the correlation between different genotypes of porcine AluI-RFLP and porcine semen quality traits was analyzed. SAS statistical software (SAS Institute Inc, Version 8.0) GLM program was used for single-marker analysis of variance, and REG program was used to calculate the additive effect and dominant effect of genes, and the significance test was carried out. The model used was:

[0036] Y ij =μ+G i +F j +e ij

[0037] Y ij is the trait phenotype value, μ is the average value, G i is ge...

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Abstract

The present invention belongs to the technical field of screening and application of pig genetic markers, and in particular relates to a genetic marker related to porcine semen quality traits and application, the genetic marker is a gene obtained by cloning of a pig RNF4 gene 3 'untranslated region, the single nucleotide polymorphism (SNP) can be used as a genetic marker of the porcine semen quality traits, a nucleotide sequence of the genetic marker is as shown in SEQ ID NO: 3 or SEQ ID NO: 4, the sequence SEQ ID NO: 3 or SEQ ID NO: 4 has an allelic gene mutation at 487bp, and AluI-RFLP polymorphism is caused by the allelic gene mutation. Screening step of the method are as follows: extraction of genomic DNA from pig blood, primer design, PCR amplification, PCR product cloning, sequencing, sequence comparing analysis, single nucleotide polymorphism detection, marking and semen quality trait correlation analysis. The genetic marker can be used in pig marker assisted selection.

Description

technical field [0001] The invention relates to the technical field of pig genetic marker screening, in particular to a molecular genetic marker related to pig semen quality traits and its application. The genetic marker of the invention obtains SNP from the 3' untranslated region of the RNF4 gene, and the invention also includes pig RNF4 Detection methods and applications of gene mutation sites. Background technique [0002] The fecundity of boars plays an important role in pig production, and the reproductive performance of boars is generally measured by identifying the quality of semen. Semen quality indicators include semen volume, sperm density, sperm morphology, and sperm motility. Genetic effect and environmental effect are two important factors affecting semen quality. Environmental factors can be artificially controlled and improved, but only by improving genetic efficiency can fundamentally improve semen quality and boar fecundity. Routine selection has been slow...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 李凤娥马昌萍李家连牛步月宋慧彬胡盼娣
Owner HUAZHONG AGRI UNIV
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