Molecular marker relative to sperm viability and total sperm number of boar and application
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A molecular marker, sperm motility technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. Sensitive effect
Active Publication Date: 2019-04-05
HUAZHONG AGRI UNIV
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However, because there is a negative correlation between some production traits and semen traits (Oh et al.Estimates of Genetic Correlations between Production and Semen Traits in Boar[J].Asian-australasian journal of animal sciences,2005,19(19):160- 164), excessive selection of boar production traits may lead to a decline in the quality of boar semen and reduce the average number of weaned piglets (PSY) provided by sows per year
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Embodiment 1
[0032] Example 1: Genotyping Detection
[0033] (1) The total DNA of Duroc boar sperm was extracted using the conventional magnetic bead method sperm genome extraction kit (Wuhan Nanomagnetic Biotechnology Co., Ltd.), the specific steps are as follows
[0034] (1) Take an appropriate amount of semen (5-15 μL) from a Duroc boar into a 1.5 mL centrifuge tube;
[0035] (2) Add 500 μL of lysate (reagents included in the above kit) and 5 μL of proteinase K (20 mg / mL) to the centrifuge tube, shake and mix for 30 seconds, then place in a 65°C oven or metal bath for 30 minutes. ~1h;
[0036] (3) After the lysis is completed, transfer all the supernatant to a deep-well plate (marked as ①), and add 350 μL of isopropanol to each well;
[0037] (4) Place the No. ① deep-well plate on station 1 of the nucleic acid extractor, and place the deep-well plates containing magnetic beads, washing solution ①, washing solution ②, washing solution ③ and eluent on station 2 respectively. ~6 on. Tu...
Embodiment 2
[0044] Example 2: Application of ALGA0001958 Molecular Marker Typing Method in Association Analysis of Duroc Boar Semen Traits
[0045] (1) Phenotypic pretreatment of semen traits of Duroc boars
[0046] The traits of boar semen belong to repeated measurement traits. It is necessary to use the mixed linear model (MLM). The joint effect of boar station, year and month is used as the fixed effect, the age in days and semen collection interval are used as covariates, and the individual is used as a random effect to calculate the individual random effect. Effects, as newly constructed phenotypes (pseudo-phenotypes) for subsequent genome-wide association analysis (GWAS). The lme4 package in the R statistical environment was used for analysis. The data included 2020 individuals and 105201 semen collection records. The specific model is as follows:
[0047]
[0048] Among them, y ijklm is the original phenotypic value of semen traits (sperm motility and total sperm count) of th...
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Abstract
The invention belongs to the technical field of screening of molecular markers of boars, and particularly relates to a molecular marker relative to sperm viability and a total sperm number of a boar and application. The molecular marker is cloned from a gene segment with a login number of ALGA 0001958. Through a gene chip technology, a gene is typed, screening is conducted to obtain the molecularmarker relative to characters of the sperm viability and the total sperm number of the boar, a marked nucleotide sequence is as shown in SEQ ID NO:1, A / G allele mutation exists in a base position of asite 101 of the sequence, and when nucleotide of the site 101 on the nucleotide sequence as shown in the SEQ ID NO:1 is A, the fact that the boar has higher sperm viability and a larger total sperm number is judged. The invention discloses a screening method of a molecular marker relative to a boar seminal fluid character and correlation analysis thereof. New SNP marker resources are provided formarker assisted selection of the characters of the sperm viability and the total sperm number of the boars.
Description
technical field [0001] The invention relates to the technical field of pig molecular marker preparation, in particular to a molecular marker related to boar sperm motility and total sperm count and its application. The molecular marker involves the use of a SNP. The molecular marker of the invention can be used for marker-assisted selection of boar sperm motility and total sperm count traits. Background technique [0002] my country is the largest producer of pigs in the world, ranking first in both the consumption of pigs and the scale of pig breeding (Liu Wentao et al. A comparative study on the pig industry in China and the United States[J]. Chinese Journal of Animal Husbandry, 2016 , 52(06):3-7). The pig raising model has gradually developed from small-scale pig farms and individual farmers to large-scale pig farms. By 2016, the proportion of large-scale pig farms and integrated pig farming enterprises has exceeded 50% (Wang Ruinian. Analysis of the status quo of large...
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