Application of mir-155 as a marker in the identification or auxiliary identification of animal semen quality

A mir-155, quality technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effect of speeding up the breeding process, high accuracy, and improving semen quality traits

Active Publication Date: 2022-05-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there are no reports on the transcriptomics of porcine SPEVs, and there are no reports on how SPEVs regulate the motility of mature sperm

Method used

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  • Application of mir-155 as a marker in the identification or auxiliary identification of animal semen quality
  • Application of mir-155 as a marker in the identification or auxiliary identification of animal semen quality
  • Application of mir-155 as a marker in the identification or auxiliary identification of animal semen quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1. Transcriptome sequencing revealed significant differences in the expression of miR-155 in large white pig seminal extracellular vesicles with high and low semen quality

[0069] 1. Experimental animals and grouping

[0070] The fresh semen samples used in this example are from Henan Jingwang Breeding Pig Improvement Co., Ltd. The boar breed is Large White pig, aged 24-36 months, and raised to sexual maturity under normal nutritional level and feeding conditions. The boars with high semen quality and the boars with low semen quality were screened according to the semen collection records of the boar stud within half a year, and 12 individuals were finally selected. All boars were divided into two groups, a high quality semen group (>90% total motility and >70% fast motility) and a low semen quality group (<80% total motility and <55% fast motility). Fresh semen samples were collected from 12 boars at the same time by semen collectors.

[0071] See Table 1 fo...

Embodiment 2

[0113] Example 2. Real-time fluorescent quantitative PCR detection of the expression level of miR-155 in the total RNA (obtained in step 3 in Example 1) of large white pig SPEVs with high and low semen quality

[0114] 1. Preparation of primer pairs

[0115] According to the nucleotide sequence of miR-155, primer pair 1 (composed of primer F1 and primer R1) for reverse transcription and primer pair 2 (composed of primer F2 and primer R2) for real-time fluorescent quantitative PCR detection were designed and synthesized. ).

[0116] The nucleotide sequences of each primer are listed in Table 3.

[0117] table 3

[0118]

[0119] 2. Acquisition of cDNA of porcine SPEVs

[0120] The total RNA of porcine SPEVs obtained in step 3 in Example 1 was respectively taken, and reverse transcription was performed using primer pair 1 to obtain cDNA of porcine SPEVs.

[0121] The reaction system was 15 μL, including 5 μL total RNA of porcine SPEVs, 0.15 μL 100 mM dNTP, 1 μL MultiScrib...

Embodiment 3

[0135] Example 3. Real-time fluorescent quantitative PCR detection of miR-155 expression in total RNA of Duroc pig SPEVs with high and low semen quality

[0136] 1. Preparation of primer pairs

[0137] Same as Step 1 in Example 2.

[0138] 2. Acquisition of cDNA of porcine SPEVs

[0139] 1. The fresh semen sample used in this example comes from Henan Jingwang Breeding Pig Improvement Co., Ltd. The boar breed is Duroc, the age is 24-36 months, and it is raised to sexual maturity under normal nutritional level and feeding conditions. The boars with high semen quality and the boars with low semen quality were screened according to the semen collection records of the boar stud within half a year, and finally 8 individuals were selected. All boars were divided into two groups, high semen quality group (total motility >90%, fast motility >70% and slow motility 15%). Fresh semen samples were collected from 8 boars at the same time by semen collectors.

[0140] See Table 5 for det...

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Abstract

The invention discloses the application of miR-155 as a marker in identifying or assisting in identifying the quality of animal semen. Detect the expression level of miR-155 in the individual SPEVs of the animal to be tested, if the expression of miR-155 in the individual SPEVs of the animal to be tested is higher than the expression of miR-155 in the individual SPEVs of the control animal, the semen quality of the animal is poor; If the expression level of miR-155 in the SPEVs of individual animals is not higher than the expression level of miR-155 in SPEVs of individual control animals, the semen quality of the animal is good; the individual control animals are animal individuals with excellent semen quality. Therefore, the quality of animal semen can be identified only by detecting the expression of miR‑155 in SPEVs of individual animals. The invention has great application value for establishing a population with excellent semen quality.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of miR-155 as a marker in identifying or assisting in identifying the quality of animal semen. Background technique [0002] RNA-seq technology, that is, transcriptome sequencing technology, is a technology for sequencing the total RNA in cells or tissues by means of next-generation high-throughput sequencing methods. RNA-seq technology can detect the expression of genes in cells or tissues, and at the same time, it can detect important genetic variations in the population through comparison with the reference genome sequence. At present, RNA-seq technology has been widely used in the study of important economic traits of animals and plants. [0003] High-quality boars are one of the most important factors in the development of pig farming. The quality of boar semen directly affects the economic benefits of pig farms. At present, artificial insemination t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/113
CPCC12Q1/6888C12Q2600/178C12Q2600/158C12Q2600/124
Inventor 姜力
Owner CHINA AGRI UNIV
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