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Molecular marker for eye muscle pH value-related gene SVEP1 within 24 hours after slaughter of pigs and application of molecular marker

A technology of SVEP1-PHU1 and SVEP1-PHU2, which is applied in the field of porcine molecular marker preparation, can solve problems such as blindness, inability to select traits, and long time-consuming breeding

Active Publication Date: 2017-04-26
HUAZHONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, most of the carcass traits and meat quality traits can only be measured by the slaughter method, and the muscle pH value also needs to be measured by the slaughter method. It is difficult to achieve great genetic progress by traditional breeding methods. Molecular marker-assisted breeding is an effective method for meat quality improvement. Tu (Liu Lu. Pork pH value-related QTL integration mapping and gene association analysis: [D]. School of Basic Medicine, Liaoning Medical University, 2012)
The traditional breeding method for the selection of muscle pH takes a long time, is blind, is easily disturbed by the external environment, and cannot accurately select traits
Since the muscle pH trait can only be measured by the slaughter method, it brings a huge workload to the breeding work

Method used

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  • Molecular marker for eye muscle pH value-related gene SVEP1 within 24 hours after slaughter of pigs and application of molecular marker
  • Molecular marker for eye muscle pH value-related gene SVEP1 within 24 hours after slaughter of pigs and application of molecular marker
  • Molecular marker for eye muscle pH value-related gene SVEP1 within 24 hours after slaughter of pigs and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The acquisition of a set of molecular markers related to the pH of the eye muscles of pigs at 24 hours after slaughter:

[0087] 1) Primer design of pig SVEP1 gene and partial DNA sequence amplification

[0088] Use the pig SVEP1 gene sequence information (GenBank accession number: ENSSSCG00000005455) as the template sequence for primer design, and use the biological primer design software Oligo7.0 to design the primers. The primer sequences are as follows:

[0089] Forward primer P1: 5’-TGCAATGTGGGGTTCCTGTT-3’;

[0090] Reverse primer F1: 5'-TGGGAAAGCAGTCCAGAGATG-3'.

[0091] Forward primer P2: 5’-ACAGGTGGCAAATGGG-3’;

[0092] Reverse primer F2: 5'-CTCGCAGACTGGATAAGG-3'.

[0093] 2) PCR amplification reaction:

[0094] TIANamp Genomic DNA Kit (purchased from Invitrogen, USA) was used to castrate an experimental population of purebred American large white pigs (the population was from Hubei Jinlin Breeding Farm) (slaughter determination and sampling were divided into 20 batches, pri...

Embodiment 2

[0102] Based on the detection method of molecular markers related to the pH value of the eye muscles of pigs 24 hours after slaughter, the steps are as follows:

[0103] 1) Establishment of sequencing diagnostic method

[0104] Forward primer P1: 5’-TGCAATGTGGGGTTCCTGTT-3’;

[0105] Reverse primer F1: 5'-TGGGAAAGCAGTCCAGAGATG-3'.

[0106] The amplified fragment length of this primer pair is 985 bp, and its sequence is shown in SEQ ID NO.1.

[0107] Establishment of PCR-RFLP diagnostic method

[0108] Forward primer P2: 5’-ACAGGTGGCAAATGGG-3’;

[0109] Reverse primer F2: 5'-CTCGCAGACTGGATAAGG-3'.

[0110] The amplified fragment length of the primer pair is 670 bp, and its sequence is shown in SEQ ID NO.2.

[0111] 2) PCR amplification conditions

[0112] The PCR reaction volume of the fragment obtained by primer pair P1 / F1 is 10μl, including 1μl of pig genomic DNA template to be tested, 0.2nmol / μl of forward and reverse primers, 5μl of PCRmix, and finally deionized water to a total volume of ...

Embodiment 3

[0122] The application of a molecular marker related to the pH value of eye muscles 24 hours after slaughter in the detection of polymorphism in different pig populations, the steps are:

[0123] The experimental population of purebred American large white castrated pigs (the population comes from Hubei Jinlin breeding farm) (174 American purebred large white castrated boar populations) was detected by PCR sequencing. The genotype and gene frequency of the mutation site in the experimental population are shown in Table 1-1. The test results show that there are three genotypes of the SVEP1 gene in the experimental population. Among them, there are 49 individuals of type AA and individuals of type AG. There are 90 individuals and 35 individuals of the GG type. The detection results of the population sequencing typing are consistent with the sequencing results, and the detection method of the present invention is reliable. This result indicates that SVEP1 gene is dominant in allele ...

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Abstract

The invention discloses a molecular marker for an eye muscle pH value-related gene SVEP1 within 24 hours after slaughter of pigs and application of the molecular marker. The molecule of the eye muscle pH value-related gene SVEP1 is marked as a group of molecular markers, comprising a molecular marker SVEP1-PHU1 and a molecular marker SVEP1-PHU2, respectively, wherein the nucleotide sequence of SVEP1-PHU1 has an SNP site A / G at 641bp; the nucleotide sequence of SVEP1-PHU2 has an SNP site C / G at 94bp. According to the invention, the group of molecular markers can be combined to rapidly predict the difference between eye muscle pH values within 24 hours after slaughter of different genotype pigs; meanwhile, the molecular marker can be used as a reliable marker of eye muscle pH value within 24 hours after slaughter in pig breeding work, effectively shortens breeding period, accelerates selection process, and promotes cultivation of new varieties.

Description

Technical field [0001] The invention relates to the field of preparation of pig molecular markers, in particular to the molecular marker of the ocular muscle pH-related gene SVEP1 24 hours after pig slaughter and its application. Background technique [0002] Since the reform and opening up, my country's animal husbandry development has achieved world-renowned achievements. The scale of animal husbandry production continues to expand, the total amount of animal products has increased substantially, and the quality of animal products has continued to improve. Especially in recent years, with the implementation of the policy of strengthening agriculture and benefiting agriculture, the development of animal husbandry has accelerated, the production method of animal husbandry has undergone positive changes, and the pace of scale, standardization, industrialization and regionalization has accelerated. Pork is the main source of meat in China, and the pig breeding industry occupies a ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/124C12Q2600/156
Inventor 李长春罗文哲李菁璇邹成张皓源胡岸赵书红余梅李新云
Owner HUAZHONG AGRICULTURAL UNIVERSITY
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