Genetic marker for character of litter size of pig utilizing WIF1 gene

A technology of genetic markers and litter size, which is applied in the determination/inspection of microorganisms, DNA preparation, DNA/RNA fragments, etc., which can solve the problems of slow progress in reproductive traits and low heritability of sows

Inactive Publication Date: 2013-12-04
HUAZHONG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The reproductive traits of sows are one of the important economic traits of pigs, but due to their low heritability and limited traits, the progress of routine selection of sows' reproductive traits has been slow

Method used

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  • Genetic marker for character of litter size of pig utilizing WIF1 gene
  • Genetic marker for character of litter size of pig utilizing WIF1 gene
  • Genetic marker for character of litter size of pig utilizing WIF1 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: The acquisition of pig WIF1 gene fragments and the establishment of polymorphism detection methods

[0028] The porcine WIF1 gene sequence was downloaded from the Ensembl database as the target sequence, and primers were designed online using Primer2 software. The primer sequences are shown in SEQ ID NO: 8 and SEQ ID NO: 9 in the sequence table, specifically as follows: forward primer WIF1-F: GAG ACC TCT GTTCAA AGC GTAA, WIF1-R: ATT TGT TGG GTT CGT GGC. The amplified region includes the 592bp genomic nucleotide sequence of the 5th intron of the pig WIF1 gene, such as figure 1 shown. 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 Taq DNA polymerase (Biostar International, Canada), and deionized water was added to a total volume of 25 μl ;PCR reaction program: pre-denaturation at 94°C for 4min; then denaturation at 94°C for 50s, annealin...

Embodiment 2

[0030] Example 2: Polymorphic distribution of genetic markers prepared by the present invention in different pig herds

[0031] The method of extracting pig genomic DNA (see Table 1 for samples) was carried out according to the method introduced by Xiong Yuan in "Introduction to Pig Biochemistry and Molecular Genetic Experiments", China Agricultural Press, 1999.

[0032] The PCR-AseI-RFLP polymorphism of pig WIF1 gene was detected in 3 groups of pigs, including 2 foreign blood-related pig groups and 1 synthetic line of Chinese lean pig new strain DIV line. The detection results are shown in Table 1. The results showed that there were 3 genotypes in DIV of large white pigs and Hubei white pigs; in all three populations tested, the A allele frequency was 0.70-1.00, and the A allele was the dominant allele (see Table 1) .

[0033] Table 1 Distribution results of pig WIF1 gene PCR-AseI-RFLP in different pig breeds

[0034]

[0035]

[0036] Note in Table 1: The above-menti...

Embodiment 3

[0037] Example 3: Association analysis of genetic markers prepared by the present invention and litter size traits

[0038] In order to determine whether the pig WIF1 gene PCR-AseI-RFLP is related to the difference in pig litter size, 110 new Chinese lean pig strains DIV and Hubei Academy of Agricultural Sciences Animal Husbandry were selected. 162 Large White pigs from the Institute of Veterinary Medicine were used as test materials. The AseI-RFLP method established in Example 2 was used to detect polymorphisms, and the correlation between different genotypes of pig AseI-RFLP and pig litter size traits was analyzed. The SAS statistical software (SAS Institute Inc, Version 8.0) GLM program was used for single-marker analysis of variance, and the 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:

[0039] Y ijk =μ+G i +Y i +P k +e ijk

[0040] Y ij is the trait phenotype value...

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Abstract

A genetic marker for character of litter size of a pig utilizing WIF1 gene belongs to the technical field of genetic markers, and relates to the technical field of detection to single nucleotide polymorphism (SNP) detection of the fifth intron of the encoding gene of a pig gene WIF1. A method for screening the genetic marker for character of litter size of the pig comprises the following steps: obtaining a genome DNA from pig blood, designing a primer, performing PCR amplification, cloning the PCR products, measuring sequence, comparing and analyzing the sequence, performing SNP detection, marking, conducting correlation analysis among the characters of litter sizes. The DNA sequence of the fifth intron of the pig gene WIF1 and the SNP genotyping detection and a genetic marker are disclosed; the nucleotide sequence is shown in SEQ ID NO: 1-7; an allele mutation exists in the 133 bp position in the sequence table of SEQ ID NO:1-7, and leads to the polymorphism of Ase1-RFLP. The invention further discloses a manufacturing method and application of the genetic marker.

Description

technical field [0001] The invention relates to the technical field of preparation of pig genetic markers, in particular to a WIF1 gene as a molecular marker for pig litter size traits and its application, which includes a detection method and application of pig WIF1 gene mutation sites. Background technique [0002] The reproductive traits of sows are one of the important economic traits of pigs, but due to their low heritability and limited traits, the progress of conventional selection of sows' reproductive traits has been slow. With the rise and rapid development of molecular biology techniques, a combination of conventional breeding and marker-assisted selection (MAS) has been developed, which can effectively accelerate the progress of selection for litter size traits in pigs. At present, the methods of molecular marker identification mainly include genome scanning method and candidate gene method. Differentially expressed genes can be studied as candidate genes if the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 李凤娥花伦王磊梅书棋陶虎彭先文雷彬张旭英杨嘉豪
Owner HUAZHONG AGRI UNIV
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