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SNP molecular marker used for screening and/detecting abnormal rate of breeding bull sperms

A technology for sperm deformity and bulls, applied in the field of molecular genetic biology, can solve problems such as unreported functional verification, and achieve the effects of increasing the economic benefits of ranches, reducing costs, and reducing blindness.

Active Publication Date: 2018-11-16
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

However, in the Chinese Holstein bull population, whether there are SNPs loci related to bull semen quality in the PRKCB gene and its functional verification have not been reported.

Method used

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  • SNP molecular marker used for screening and/detecting abnormal rate of breeding bull sperms
  • SNP molecular marker used for screening and/detecting abnormal rate of breeding bull sperms
  • SNP molecular marker used for screening and/detecting abnormal rate of breeding bull sperms

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Identification of the SNP site and genotype combination of embodiment 1 bovine PRKCB gene

[0032] The invention adopts the PCR product direct sequencing method to carry out SNP identification and typing on the PRKCB gene of Chinese Holstein bulls.

[0033] 1.1 Experimental materials

[0034] The bull used in the present invention is selected from Shandong Oakes bull station, 117 Chinese Holstein bulls. Frozen bull semen was stored in liquid nitrogen for later use.

[0035] 1.2 Extraction of genomic DNA from frozen semen

[0036] 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.

[0037] 1.3 Design of primers

[0038] According to the bovine PRKCB gene sequence (accession number: AC_000182) provided by NCBI, utilize Primer Premier5.0 software to design primers, the se...

Embodiment 2

[0050] Example 2 Correlation Analysis of Bovine PRKCB Gene Haplotype Combination and Bull Sperm Motility

[0051] SAS 9.0 was used to compare the correlation between different genotype combinations of PRKCB gene and sperm abnormality rate in Chinese Holstein bulls. Its model is:

[0052] Y ijk =μ+G i +H k +e ijk

[0053] Y ijk is the observed value of the trait of sperm deformity in bulls; μ is the average value of the population; G i : fixed effect of haplotype combination; H k : fixed effect of the number of sessions; e ijk : random residual effect.

[0054] The results of haplotype combination correlation showed that among the six genotype combinations, the sperm abnormality rate of bulls with H1H2 haplotype combination was significantly lower than that of bulls with H1H1 and H2H2 haplotype combination (P<0.05) (Table 1).

[0055] Table 1 Correlation analysis between different haplotype combinations of bovine PRKCB gene and semen quality

[0056]

[0057] Note...

Embodiment 3

[0060] Embodiment 3 detection kit

[0061] As described in Example 1, bovine PRKCB genes g.-1526C>T, g.-1409C>T, g.-1259C>T, g.-1249A>G, g.-1230C>T, g.-1220C> T, g.-1091C>T, g.-1089C>T, g.-1088C>T, g.-1081A>T, g.-869A>C and g.-806A>C are all related to bull sperm deformity rate are closely related. Therefore, it is possible to invent a PRKCB gene SNPs molecular marker detection kit suitable for screening the abnormality rate of male bulls.

[0062] 1.1 PCR-RFLP genotyping

[0063] Since the 12 SNPs in the PRKCB promoter region are completely linked, the genotype of the overall SNPs can be obtained by detecting the genotype of any one of the SNPs. Here, we take the g.-1259C>T site as the detection point, as long as the g.-1259C>T site genotype is detected, the genotypes of the remaining 11 SNPs sites can be known. In order to find a simpler method, we use the PCR-RFLP method to identify the genotype of one of the SNPs, and then we can know the genotype of the entire SNPs co...

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Abstract

The invention belongs to the technical field of molecular genetics, and concretely relates to a SNP molecular marker used for screening and / detecting abnormal rate of breeding bull sperms. A PRKCB gene comprises 12 completely linked SNP sites, 2 haplotypes can be obtained through haplotype analysis, which are H1 (TCTGTTTCTAAC) and H2 (CTCACCCTCTCA), and three haplotype combination H1H1, H1H2, andH2H2 are provided. The association analysis with the bull seminal fluid quality shows that the abnormal rate of the H1H2 haplotype combination bull frozen semen after unfreezing is obviously lower than that of H1H1 and H2H2 haplotype combination bull (P is less than 0.05).

Description

technical field [0001] The invention belongs to the technical field of molecular genetics, and in particular relates to a group of SNP molecular markers used for screening and / or detecting the abnormality rate of sperm of breeding bulls. Background technique [0002] The selection of excellent breeding bulls plays a decisive role in the genetic improvement and reproduction of dairy cows, and is of great significance to the sustainable and healthy development of the modern dairy industry. According to statistics, breeding bulls contribute more than 75% to the genetic progress of dairy cow populations. The quality of semen is directly related to whether the cow is pregnant or not. At present, in the developed countries of the dairy industry, an excellent breeding bull can produce 80,000 to 100,000 doses of frozen semen per year, up to 250,000 doses, which is significantly higher than the average annual yield of 21,800 doses of bulls in my country. The quality of bull semen i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 杨春红王长法孙艳王金鹏高琦璨鞠志花黄金明王秀革姜强
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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