A set of SNP molecular markers for screening and/or detecting ejaculatory volume and fresh sperm motility of breeding bulls

A technology for sperm motility and bulls, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of ejaculation volume and fresh semen vitality of breeding bulls, so as to increase the economic benefits of pastures and is widely used , high accuracy effect

Active Publication Date: 2019-03-15
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is still no report on the SNPs of the INCENP gene and the genotype combination of the INCENP gene that affects the amount of ejaculation and fresh sperm vitality of the bulls in the prior art.

Method used

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  • A set of SNP molecular markers for screening and/or detecting ejaculatory volume and fresh sperm motility of breeding bulls
  • A set of SNP molecular markers for screening and/or detecting ejaculatory volume and fresh sperm motility of breeding bulls
  • A set of SNP molecular markers for screening and/or detecting ejaculatory volume and fresh sperm motility of breeding bulls

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The SNP locus of embodiment 1 bovine INCENP gene and the identification of genotype combination

[0039] The invention adopts the means of direct sequencing and PCR-RFLP to carry out SNP identification on bovine INCENP.

[0040] 1.1 Experimental materials

[0041] The bulls used in the present invention are selected from three domestic bull breeding stations: Beijing Dairy Cow Center (83 heads), Shanghai Guangming Holstein Animal Husbandry Co., Ltd. (56 heads), and Shandong Oakes Bull Breeding Station (125 heads). Frozen bull semen was stored in liquid nitrogen for later use.

[0042] 1.2 Extraction of genomic DNA from frozen semen

[0043] The high-salt method was used to extract the genomic DNA from the frozen semen, and the purity and concentration of the DNA samples were detected by a nucleic acid and protein analyzer, and finally diluted to 50 ng / μl, and stored at 4°C for later use.

[0044] 1.3 Design of primers

[0045] According to the bovine INCENP gene seq...

Embodiment 2

[0063] Example 2 Association Analysis of Cattle INCENP Gene Haplotype Combination and Bull Sperm Motility

[0064] SAS 9.0 was used to compare the correlation of different genotype combinations of INCENP gene with ejaculate volume and fresh semen motility. Its model is:

[0065] Y ijk =μ+G i +Y j +H k +e ijk

[0066] Y ijk is the observed value of ejaculation volume or sperm motility traits of bulls; μ is the average value of the population; G i : fixed effect of haplotype combination; Y j : fixed effect of season; H k : fixed effect of the number of sessions; e ijk : random residual effect.

[0067] The results of haplotype combination association showed that among the eight genotype combinations, the ejaculation volume of H3H4 genotype bulls was significantly higher than that of H1H4 genotype bulls (P<0.05), and was extremely significantly higher than that of H1H3, H2H4, H3H3 genotype individuals Bulls (P<0.01); H3H4 genotype combination bulls had significantly h...

Embodiment 3

[0072] Embodiment 3 detection kit

[0073] As described in Example 1, A>G at the 19970th position and T>G at the 34378th position of the bovine INCENP gene are closely related to the ejaculatory volume and fresh semen vigor of the bull. Therefore, it is possible to invent an INCENP gene SNP molecular marker detection kit suitable for screening bull ejaculation volume and fresh sperm vigor, that is, design specific primers for bovine INCENP gene according to these two SNP sites for amplification and detection.

[0074] Prepare a test kit for the detection of bull sperm motility, and the detailed components are shown in Table 2:

[0075] Table 2 The components of the test kits for the detection of bull sperm motility

[0076]

[0077] We first collected the semen of the breeding bulls, stored them in liquid nitrogen for later use, and extracted the total DNA from the frozen semen using the high-salt method. The components in the above kit were used to perform PCR amplificatio...

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Abstract

The invention relates to a group of SNP (single nucleotide polymorphism) molecular markers for screening and detecting the ejaculatory amount and the fresh sperm activity level of stud bulls. Specifically, SNPs sites are positioned at the 19970th and 34378th basic groups of INCENP genes, the two SNPs sites have multiple types of genotype combinations in stud bull groups, and the genotype combinations of the INCENP genes of frozen sperms of the stud bulls are detected and the ejaculatory amount and the fresh sperm activity of the stud bulls are predicted by extracting the genome DNA of the frozen sperms of the stud bulls. The SNPs of the INCENP genes of the stud bulls are successfully screened out, the correlation between the mononucleotide polymorphism of the INCENP genes of the stud bulls and the ejaculatory amount and the fresh sperm activity of the stud bulls is illuminated for the first time, a simple, convenient and efficient method for detecting the ejaculatory amount and the fresh sperm activity of the stud bulls is provided, and a corresponding detection kit is developed. The kit has the advantages of low cost, stable reliability, high efficiency and the like, and can be used for screening early high-quality stud bulls.

Description

technical field [0001] The present invention relates to the technical field of livestock molecular genetics, in particular to a set of SNP molecular markers for screening and / or detecting ejaculatory volume and fresh semen vigor of breeding bulls, and in particular to detection of ejaculatory volume and fresh semen activity of breeding bulls through molecular marker technology The SNPs locus of the INCENP gene related to the level of vigor provides a method and kit for detecting the amount of ejaculation and the vigor of fresh semen of bulls, and is applied to the early breeding of excellent livestock reproductive traits. Background technique [0002] Milk has high nutritional value and is an indispensable food in our daily life. Chinese Holstein cows are the breed with the widest breeding range and the largest milk yield in China. Chinese Holstein bulls are an important part of dairy cow breeding. The quality of their semen not only determines the economic benefits of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 王长法刘娟孙艳姜强杨春红刘保申鞠志花王秀革黄金明仲跻峰
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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