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Molecular marker and its application of gene svep1 related to eye muscle pH value 24 hours after slaughter

A SVEP1-PHU1, 24-hour technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of inability to select traits, susceptibility to external environment interference, long time-consuming breeding, etc.

Active Publication Date: 2019-08-06
HUAZHONG AGRI UNIV
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  • 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 and its application of gene svep1 related to eye muscle pH value 24 hours after slaughter
  • Molecular marker and its application of gene svep1 related to eye muscle pH value 24 hours after slaughter
  • Molecular marker and its application of gene svep1 related to eye muscle pH value 24 hours after slaughter

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Obtaining a set of molecular markers related to the pH value of pig eye muscle 24 hours after slaughter:

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

[0088] Using the porcine SVEP1 gene sequence information (GenBank accession number: ENSSSCG00000005455) as the template sequence for primer design, use the biological primer design software Oligo7.0 to design 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] Using the TIANamp Genomic DNA Kit kit (purchased from Invitrogen, USA) from the experimental group of purebred American large white castrated pigs (the group originated from the Hubei Jinlin Breeding Farm) (slaughter measurement and sampling were divided into 20 ...

Embodiment 2

[0102] Based on the detection method of molecular markers related to the pH value of eye muscle 24 hours after pig 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 length of the fragment amplified by the primer pair is 985bp, 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 length of the fragment amplified by the primer pair is 670bp, and its sequence is shown in SEQ ID NO.2.

[0111] 2) PCR amplification conditions

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

Embodiment 3

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

[0123] The experimental group of purebred American Large White castrated pigs (the group came from Hubei Jinlin Breeding Farm) was detected by PCR sequencing (174 American purebred Large White castrated boar groups). 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, of which there are 49 AA-type individuals and 49 AG-type individuals. 90 individuals, 35 individuals of GG type, the detection results of population sequencing and typing are consistent with the sequencing results, and the detection method of the present invention is reliable. The results indicated that allele A of SVEP1 gene was dominant in the experimental population of purebre...

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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 gene SVEP1 related to eye muscle pH value 24 hours after pig slaughter and its application. Background technique [0002] Since the reform and opening up, the development of my country's animal husbandry has made remarkable achievements. The production scale of animal husbandry has been continuously expanded, the total amount of animal products has increased significantly, 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 farmers, the development momentum of animal husbandry has accelerated, the production mode of animal husbandry has undergone positive changes, and the pace of scale, standardization, industrialization and regionalization has accelerated. Pork is the most important source of meat in China, and 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 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 AGRI UNIV
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