Molecular marker of loin-eye area character-related gene SVEP1 and application of molecular marker
A technology related to molecular markers and molecular markers, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve problems such as stress caused by measuring environmental influences, affecting breeding effects, time-consuming and labor-intensive, etc.
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Examples
Embodiment 1
[0061] Acquisition of a Molecular Marker Associated with Eye Muscle Area Traits:
[0062] 1) Primer design and partial DNA sequence amplification of porcine SVEP1 gene
[0063] 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:
[0064] Forward primer: 5'-TGTCGCATGTGCTTGGTAGAT-3';
[0065] Reverse primer: 5'-GAACCTGGACTTCAAGGGACCATTCTTCCTCTCTCAACATC-3';
[0066] 2) PCR amplification reaction:
[0067] Using the TIANamp Genomic DNA Kit kit (purchased from Invitrogen, USA) from the experimental group of purebred American large white castrated pigs (this group comes from Hubei Jinlin Breeding Farm) (slaughter measurement and sampling were divided into 21 batches, and the number of individuals was determined A total of 207 pigs, all of which were American purebred Large White castrate...
Embodiment 2
[0073] A detection method based on molecular markers related to eye muscle area traits, the steps are as follows:
[0074] Establishment of PCR-RFLP diagnostic method
[0075] 1) Primer sequence
[0076] Forward primer: 5'-TGTCGCATGTGCTTGGTAGAT-3';
[0077] Reverse primer: 5'-GAACCTGGACTTCAAGGGACCATTCTTCCTCTCTCAACATC-3';
[0078] The length of the fragment amplified by the primer is 579bp, and its sequence is shown in SEQ ID NO.2.
[0079] 2) PCR amplification conditions
[0080] The total volume of PCR reaction is 10 μl, including 1 μl of porcine genomic DNA template to be tested, 0.2 nmol / μl of forward and reverse primers, 5 μl of PCR mix, and finally adding deionized water to a total volume of 10 μl. The PCR reaction conditions were as follows: 95°C pre-denaturation for 5 minutes, followed by 31 cycles of 95°C denaturation for 30 s, 60°C annealing for 30 s, and 72°C extension for 36 s; finally, 72°C extension for 5 min; PCR products were detected by 2% agarose gel elect...
Embodiment 3
[0085] Application of a molecular marker related to eye muscle area traits in polymorphism detection in different pig populations, the steps are:
[0086] PCR-Sfan I-RFLP was used to detect the purebred American large white castrated pig experimental group (the group came from Hubei Jinlin Breeding Farm) (207 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. The test results show that there are three genotypes of the SVEP1 gene in the experimental population, of which there are 47 AA-type individuals and 99 AG-type individuals , there were 61 individuals of GG type, and the detection result of enzyme digestion typing was consistent with the sequencing result, and the detection method of the present invention was reliable. The results indicated that allele G of SVEP1 gene was dominant in the experimental population of purebred American large white castrated pigs.
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Application Information
- IPC
- C12Q1/68; C12N15/11
- CPC
- C12Q1/6888; C12Q2600/124; C12Q2600/156
- Inventors
- 李长春; 罗文哲