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Method for evaluating sperm viability of bulls based on FBXW11 methylation and application

A sperm motility and methylation technology, applied in the field of livestock molecular genetic biology, can solve problems such as technology application, and achieve the effects of improving offspring quality, convenient evaluation, and convenient use

Active Publication Date: 2019-08-20
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the genes and markers related to semen quality have been identified from the level of genetics. The related research is still in the early stage, and no mature technology has been applied to production.

Method used

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  • Method for evaluating sperm viability of bulls based on FBXW11 methylation and application
  • Method for evaluating sperm viability of bulls based on FBXW11 methylation and application
  • Method for evaluating sperm viability of bulls based on FBXW11 methylation and application

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Experimental program
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Embodiment 1

[0049] In this embodiment, the detection method for the methylation site of the FBXW11 gene is described, and the method includes:

[0050] S1. Collection and storage of semen samples;

[0051] S2. DNA extraction and concentration determination;

[0052] S3. Genomic DNA is treated with bisulfite;

[0053] S4. Nested PCR, landing PCR, and Bisulfite sequencing PCR amplify methylation sites;

[0054] S5. Alignment and identification of methylation sites.

[0055] Wherein, in step S1, the specific method includes:

[0056] S1.1 Collect the semen of the bull to be tested, record the corresponding semen quality parameters, and number the semen samples according to the production records;

[0057] S1.2 Collect the fresh semen of each bull and immediately put it into liquid nitrogen for storage.

[0058] In step S2, the method specifically includes:

[0059] S2.1 Genomic DNA is extracted from each semen sample;

[0060] S2.2 Identify the concentration and purity of the genomic ...

Embodiment 2

[0088] The semen samples of 9 full-sib paired bulls were grouped and analyzed, and the experimental groups were determined according to the sperm motility. The details are as follows: 9 Chinese Holstein bulls have complete semen quality production records. The frequency of bull semen collection is 1 week / 1 time, and the data records include: semen volume, sperm motility, sperm density, and deformity rate. According to the national standard "GB 4143-2008 Bovine Frozen Semen", the samples were divided into two groups: high sperm motility group (≥65%) and low sperm motility group (<60%), as shown in Table 2, where H1-H4 are high The vitality group, L1-L5 is the low vitality group.

[0089] Table 2 Production records of semen quality of 9 breeding bulls

[0090]

[0091]

[0092] The sequencing results of the above nine groups of semen samples analyzed by fluorescent real-time quantitative PCR technology showed that there were differences in the expression of FBXW11 in the...

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Abstract

The invention belongs to the technical field of livestock molecule heredity biology, and particularly relates to a method for evaluating sperm viability of bulls based on FBXW11 methylation and an application. High-flux sequencing is performed on a bull semen transcriptome. A FBXW11 gene and 11 methylation sites influencing the sperm viability of the bulls are provided, through functional annotation and data confluence analysis, a methylation site mark for evaluating the degree of the sperm viability of the bulls is developed, and a Bisulfite sequencing PCR detection method for several key methylation sites is established and can be used for evaluating the sperm viability of breeding bulls. Through the adoption of the technical scheme of the invention, the DNA methylation difference situation of the gene is analyzed, so that individuals or semen samples carrying ideal properties are screened, bulls having high fecundity and semen having high sperm viability are bred out, the propagation efficiency of the breeding bulls and cows is improved, and the purpose of increasing the economic benefits is achieved.

Description

technical field [0001] The disclosure belongs to the field of livestock molecular genetic biology technology, and specifically relates to a method for evaluating the motility of bull sperm by detecting the methylation rate of 11 sites in the FBXW11 gene sequence and the application of corresponding reagents and primers in the preparation of kits . Background technique [0002] The information disclosed in this Background section is only intended to increase the understanding of the general background of the disclosure, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes prior art that is already known to those skilled in the art. [0003] Optimizing reproductive efficiency is an important way to improve the production efficiency and economic benefits of dairy cows. Artificial insemination technology is the only breeding technology that has been widely and successfully applied in the dairy industry so far, and ha...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888G01N33/573A01K67/02
CPCC12Q1/6888G01N33/573A01K67/02C12Q2600/154C12Q2600/124C12Q2600/158G01N2333/9015
Inventor 黄金明刘勇魏晓超王秀革鞠志花姜强李彦芹赵晗高亚平刘文浩张亚冉王金鹏杨春红孙艳王玲玲
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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