KR101341813B1 - SNP Markers for identification of pig and diagnosing method using the same - Google Patents

Abstract

The present invention relates to a monobasic polymorphic gene marker and diagnostic method for individual identification and traceability of pigs. By utilizing the marker factor according to the present invention, it is possible to measure the genetic ability more accurately than the selection effect by the phenotype. In addition, it is possible to use as a genetic analysis technique for pork history management, and it is possible to improve the reliability of the analysis because it is possible to combine the functional analysis tools such as RFID and genetic analysis method.

Description

SNP Markers for identification of pig and diagnosing method using the same}

The present invention relates to a single nucleotide polymorphism gene marker for pig identification and history management and pig identification method using the same.

In selecting the vertical axis, the selection was based on the selection index formula based on the phenotype. Recently, the selection or selection of variants using molecular markers is not only a trait controlled by a single gene, but also a marker-assisted selection of traits controlled by Quantitative Trait Loci (QTL). Simulations show that MAS can be effectively used. In general, molecular markers can be used to measure genetic capacity 25-35% more accurately than the selection effect by phenotypic value, which can dramatically increase heritability. In the case of pig hog and piglet selection, the introduction of DNA markers in the existing method based on phenotypic value can improve the accuracy of selection, thereby maximizing genetic improvement.

The genes or markers currently used in the hog industry are HAL (flesh), ESR (liver number), PRLR (liver number), RBP4 (liver number), KIT (white hair color), MC1R (red / black hair color), MC4R ( Growth, obesity), FUT1 (edema disease), RN (flesh), AFABP (myopathy), HFABP (myopathy), PRKAG3 (flesh), IGF2 (conductor composition), and the like.

Seed income is estimated at 1,800 heads (1,836 heads per year), which is about 4 billion won a year, and about 30% replacement (550 heads) is expected to save about 1 billion won annually.

In recent years, the inclusion of genomic information in breeders in cattle and chickens is a global trend. Pig-related monobasic polymorphisms will be very effective as genetic DNA markers for pig selection.

In order to promote the planned pork tracer system, it is necessary to develop DNA markers that can be used for individual identification and traceability in the pig production slaughter distribution stage.

Prior arts in this field include Korean Patent Publication No. 2011-0067941 'Marker Factor for Identifying Increased Muscle Fiber Type in Pigs' and US Patent 7435543 'Genetic Markers for Pig Backfat Thickness'.

Under these technical backgrounds, the present inventors have made efforts to develop a monobasic polymorphic gene marker and a diagnostic method for individual identification and traceability of pigs.

After all, an object of the present invention is to provide a polynucleotide for the identification and traceability of pigs of SEQ ID NO: 1 to 38.

Still another object of the present invention is to provide a microarray for detecting monobasic polymorphism comprising the polynucleotide.

Still another object of the present invention is to provide a kit for detecting monobasic polymorphism comprising the polynucleotide.

Still another object of the present invention is to provide a diagnostic method for individual identification and traceability of swine using the polynucleotide.

According to an aspect of the present invention, a pig individual identification method may be provided including identifying a single nucleotide polymorphism, which is the 61 st nucleotide sequence of one or more polynucleotides selected from the group consisting of SEQ ID NOs: 1 to 38.

According to one embodiment, the step of identifying the monobasic polymorphism is allele-specific probe hybridization (allele-specific probe hybridization), allele-specific amplification (allele-specific amplification), sequencing (sequencing), 5 5 'nuclease digestion, molecular beacon assay, oligonucleotide ligation assay, size analysis and single-stranded polymorphism stranded conformation polymorphism).

According to another aspect of the present invention, a polynucleotide for recognizing the nucleotide sequence of the single nucleotide polymorphism, a polypeptide encoded by it, or a microarray for detecting a single nucleotide polymorphism comprising the cDNA of the polynucleotide may be provided.

According to another aspect of the present invention, a kit for detecting a single nucleotide polymorphism comprising a polynucleotide for recognizing a nucleotide sequence of the single nucleotide polymorphism, a polypeptide encoded therein or a cDNA of the polynucleotide may be provided.

By utilizing the marker factor according to the present invention, it is possible to measure the genetic ability more accurately than the selection effect by the phenotype. In addition, it is possible to use as a genetic analysis technique for pork history management, and it is possible to improve the reliability of the analysis because it is possible to combine the functional analysis tools such as RFID and genetic analysis method.

Hereinafter, the present invention will be described in more detail.

According to an aspect of the present invention, a pig individual identification method may be provided including identifying a single nucleotide polymorphism, which is the 61 st nucleotide sequence of one or more polynucleotides selected from the group consisting of SEQ ID NOs: 1 to 38.

SEQ ID NO: Allele A Allele a One T C 2 A G 3 T C 4 T G 5 A G 6 A G 7 T C 8 T C 9 A G 10 T C 11 T G 12 T C 13 A G 14 A G 15 T C 16 A G 17 T G 18 T C 19 T C 20 A G 21 A G 22 A G 23 T C 24 T C 25 T G 26 T C 27 A G 28 T C 29 T C 30 A G 31 T C 32 A G 33 A C 34 A G 35 A G 36 T C 37 A C 38 T C

The locus of the monobasic polymorph is selected by analyzing the individual identification rate.

Individuals of pigs can be identified by checking the nucleotide sequence of the single nucleotide polymorphism.

Since there are a total of 38 single nucleotide polymorphisms, it is possible to identify 2 ³⁸ individuals in theory, and in terms of identification and history tracking for each individual pig, considering that there are about 10 million to 15 million pigs produced in Korea in general, That's enough.

Since the single base polymorphism can be confirmed even after slaughter, if a database of the single base polymorphism for the pigs being raised in each pig farm is secured, it can be used for historical tracking for various purposes.

There is no particular limitation on the method for identifying the single base polymorph, and a method generally used in the art may be used.

According to one embodiment, the step of identifying the monobasic polymorphism is allele-specific probe hybridization (allele-specific probe hybridization), allele-specific amplification (allele-specific amplification), sequencing (sequencing), 5 5 'nuclease digestion, molecular beacon assay, oligonucleotide ligation assay, size analysis and single-stranded polymorphism stranded conformation polymorphism).

According to another aspect of the present invention, a polynucleotide for recognizing the nucleotide sequence of the single nucleotide polymorphism, a polypeptide encoded by it, or a microarray for detecting a single nucleotide polymorphism comprising the cDNA of the polynucleotide may be provided.

According to another aspect of the present invention, a kit for detecting a single nucleotide polymorphism comprising a polynucleotide for recognizing a nucleotide sequence of the single nucleotide polymorphism, a polypeptide encoded therein or a cDNA of the polynucleotide may be provided.

Hereinafter, the present invention will be described in more detail with reference to Examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention.

Example  One. For five pig breeds SNP Marker  Selection and frequency analysis

Using Porcine 60K SNP chip test results for 200 pigs, including 25 conventional pigs, Duroc 51, Berkshire 28, Landrace 47, and 49 Yorkshire, 205 SNPs with a gene frequency of 0.450.55 were selected first.

Of the 205 SNPs selected as the first, except for two land races with low analysis rate, 198 dogs were considered on the chromosome and genotyping rate of the SNP locus, and the individual identification rate of each locus combination was analyzed using the cervus program. 38 SNPs were selected.

Example  2. Porcine  60K SNP chip Mass using SNP  analysis

DNA was extracted from the blood of 551 pigs containing meat-like data using Wizard Genomic DNA Purification Kit (Promega, Madison, WI, USA). SNP genotyping was performed using (Illumina, San Diego, CA, USA).

<Day 1- Amplification >

Experimental Method

1. Dispense 20ul of MA1 onto a MIDI plate (labeled MSA3 plate forward) with the MSA3 barcode.

2. Insert 4ul of DNA into MSA3 plate.

3. Write the DNA ID and location of the MSA3 plate on the Lab tracking form.

4. Add 4ul of 0.1N NaOH to each well of MSA3 plate containing MA1 and DNA.

5. Cover the MSA3 plate using a 96well cap mat and vortex for 1 minute at 1600 rpm.

6. Centrifuge at 280g for 1 minute.

7. React at room temperature for 10 minutes.

8. Add 34 ul of MA2 to each well of the MSA3 plate containing the sample.

9. Add 38 ul of MSM to each well of the MSA3 plate containing the sample.

10. Cover the cap mat and centrifuge at 280g for 1 minute.

11. React in 37 Illumina Hybridization ovens for 20-24 hours. (Amplification)

<Day 2 of experiment- Fragment >

Experimental Method

1. Remove the plate from the oven and centrifuge for 1 minute at 50 g.

2. Add 25 ul of FMS to each well containing the sample.

3. Cover the MSA3 plate with a cap mat and vortex at 1600 rpm for 1 minute.

4. Remove the plate and centrifuge at 50g for 1 minute.

5. React in 37 heat block for 1 hour.

<Day 2- Precipitation >

Experimental Method

1. Remove the cap mat and add 25ul of PM1 to each well containing the sample.

2. Cover the cap mat and centrifuge for 1 minute at 1600rpm.

3. The reaction is carried out at 37 ° C. for 5 minutes.

4. Remove the plate and centrifuge at 50g for 1 minute.

5. Remove the cap mat and add 155ul of 2-propanol to each well containing the sample.

6. Cover the plate with the new cap mat, invert 10 times, mix and store for 4 to 30 minutes.

7. Remove the MSA3 plate from the centrifuge immediately after centrifugation at 4 ° C and 3000 rpm for 20 minutes.

8. Remove the cap mat immediately and turn it over quickly to discard the supernatant.

9. Tap on absorbent pads (kitchen towel, kim towel, etc.) about 10 times.

10. Place the inverted plate on the tube rack and let it dry for 1 hour.

<Day 2- Resuspend >

Experimental Method

1. Add 23 ul of RA1 to each well containing DNA pellet and store the remaining RA1 for XStain HD Bead Chips (frozen storage).

2. Place the foil seal on the MSA3 plate and seal it by pressing the heat-sealer block for 5 seconds.

3. React for 1 hour in the Illumina Hybridization oven at 48 ℃.

4. Vortex for 1 minute at 1800 rpm.

5. Centrifuge at 280g for 1 minute.

<Day 2- Hybridization >

Experimental Method

1.Denature MSA3 plate for 20 minutes in a 95 ℃ heat block.

2. After 20 minutes, remove the MSA3 plate from the heat block and allow it to cool at room temperature for 30 minutes.

3. Insert HybChamber Gaskets into HybChamber when the plate is almost 30 minutes to cool down.

4. Put 400ul of PB2 into 8 humidifying buffer reservoirs in HybChamber and close HybChamber lid at room temperature.

5. After cooling the DNA at room temperature for 30 minutes, centrifuge 280g for MSA3 plate for 1 minute.

6. Remove the chips from the refrigerator one by one, remove the chip guarantee, align the barcode shape of the HybChamber insert with the barcode part of the chips, and load the sample into both parts of the chips using 15 ul per sample using a multichannel pipette.

7. As soon as sample loading of each chip is finished, put into HybChamber and repeat the following chips in the same way.

8. Once the chamber is filled, close the chamber lid and place it in the 48 Illumina Hybridization oven to set a speed of 5 to react for 16-24 hours.

<Day 3- Washing bead chips >

Experimental Method

1. Remove the Hyb chamber from the Hybridization oven.

2. Open the lock on the Hyb chamber and remove one insert at a time from the chamber.

3. Pull out the seal attached to the chip and remove it from the chip.

4. Place the chip without the seal into the Wash Rack and place it in the WB1 Wash dish.

5. Once all chips are in WB1, wash the rack by removing it from the dish for 1 minute and move the wash rack to another wash dish containing PB1. Repeat this process for 1 minute.

6. After dipping in PB1 wash dish, wash the rack by removing it from the dish for 1 minute, and then move the wash rack to another wash dish containing PB1 and repeat this process for 1 minute.

7. After the washing is finished, place the back frame on the BeadChips Alignment fixture, place the chips one by one in the direction of the barcode, and fit the white and separated transparent spaces to the top and bottom of the alignment fixture.

8. Raise the space, raise the Alignmet bar on the top of the chip (the area without the bar code), cover the glass plate so that the end of the glass plate sticks to the bar, and then clip it.

   (Flow-through chamber assembly completed)

9. After inserting the clip, remove the alignment bar and use scissors to cut the space at both ends of the flow-through chamber assembly.

<Day 3- XStain Beadchips >

Experimental Method

1. When the temperature of the chamber rack reaches 44 ℃, insert the flow-through chamber assembly into the chamber rack.

2. Add 150ul of RA1 to each chip and react for 30 seconds. Repeat this process five more times.

3. Add 450ul of XC1 to each chip and let it react for 10 minutes.

4. Add 450ul of XC2 to each chip and let it react for 10 minutes.

5. Add 200ul of TEM to each chip and let it react for 10 minutes.

6. Add 450ul of 95% formamide / 1mM EDTA to each chip and let it react for 1 minute.

7. React for 5 minutes.

8. Check the temperature on the label of the LTM tube and change the temperature of the chamber rack accordingly.

9. Add 450ul of XC3 to each chip, react for 1 minute, put it in again, and wait until it reaches 8 times.

10. Add 250ul of LTM to each chip and let it react for 10 minutes.

11. Add 450ul of XC3, add one more time and let it react for 5 minutes.

12. Add 250ul of ATM to each chip and let it react for 10 minutes.

13. Add 450ul of XC3, and after one minute, add one more time and let it react for 5 minutes.

14. Add 250ul of LTM to each chip and let it react for 10 minutes.

15. Add 450ul of XC3, add one more time and let it react for 5 minutes.

16. Add 250ul of ATM to each chip and let it react for 10 minutes.

17 Add 450ul of XC3, add 1 minute later, and then react for 5 minutes.

18. Add 250ul of LTM to each chip and let it react for 10 minutes.

19. Add 450ul of XC3, add one more time and let it react for 5 minutes.

20. At the end of this procedure, immediately remove the chamber rack from the flow-through chamber, transfer it to the room temperature test table, and remove it flat.

21. Add 310 ml of PB1 to the wash container and place the dye rack into the container.

22. Remove the clip from the chamber rack using the instrument, lift off the glass block, and remove the spaces on both ends of the chips so that the bead portion of the chips cannot be touched.

23. After removing all the deposits on the chip, bloom in the staining rack contained in PB1 and soak in PB1. Treat all chips in the same way.

24. Slowly move the dyeing rack up and down 10 times to quench the chips and soak for 5 minutes.

25. Fill 310 ml of XC4 in another cleaning container, quench for 10 times in the same manner as in No. 24, and soak for 5 minutes.

26. After 5 minutes, remove the dyeing rack from the cleaning container and place it on the tube rack as shown below.

27. Using the forceps, carefully remove the chips from the rack and place them on the tube rack.

28. Carefully place the tube rack with the chips in a vacuum dryer and dry for 55-55 minutes in a vacuum of 508 mmHG (0.68 bar).

29. If the chips are found to be dry, close the edges of the chips with a Kimwipe soaked in ethanol. Be careful not to touch the bead part.

30. Beadchips should be imaged using a scanner within 72 hours of completion of the experiment.

The table below relates to sequencing information of SNPs for swine identification and traceability.

number SNP name Base sequence One ALGA0002500 AGTAAAGAGCAACCGTTTCCATGTGTGCGAAATTTTACAACACATTTTCAAGGCACACAT [T / C] GTGCACAATCTAAATGACATTAAAGCTGATTGTGCATGCGTGCGTGTGTATACAGAGGCA 2 ALGA0003632 GCTGCTTTTAGAATCCTTGCTTTAACTTTTGCCATTTTTATTGTAATATGTCTTGGTGTG [A / G] GTCTGTTTGGCCTCACCTTGATTGGAGCTCTCTGTGCTTGCTGTATCTTGATGTCTGTTT 3 ALGA0005188 ACTAGTTAAGCAAGCCTGCCTTGTTCTGAGGGCTTTCCTGAACTTCCAGTGGCCTCTCAG [T / C] ATCCCCCAGCTTTCCCTCTTTATCCGTGATCCCCTATTGGTGCTTCTACAACCACCTGTG 4 ALGA0020170 CAGAGGGGCCCCCACGCTGACGGGTGACTCCCCATTAAAAGCTGTCTGATCAGGTCATGG [T / G] CTGGGAAGGGAGGGAACGGATCTGGCTGGCACAAGGGGCTACTCAGTTGAGTCTGGCCTC 5 ALGA0028052 AGAAAAGCAACAAAAAATTACTCAAAATTATTTCAGACTTTTAAAAAGGCCCCTGAAGCT [A / G] GCCATGTTCTGTGCTCCTCCTTGTTCCTTTAAGTGTACACAGACAGGGAGTTCCCATCAT 6 ALGA0033986 CCTCAGAAAACAGGATGTAAGTTGGTTGGTGTCACATCTTGGTTAGGATAAGCAGCTGCT [A / G] TTGATAGCAACTGATTAGTTCTAAGGACTGGTTATATACCACACGTGAACTTCAAAAGTG 7 ALGA0037105 TGCCGCCCCCTGTTCTCCCTCCAAGTTCAAAACACTAAGAAGAGCTGTATGAAGTCCAGT [T / C] GGCCTCTGGACCAGTTGCCAACAGGATCTTGTAAGGAAATCTGACAAAGGGTCAGATCTT 8 ALGA0043483 TTAGCCACAGGGTAGGAAAAGAATCTATCTCTCAGTCTTTTCCAATAAACCATAAGCTGC [T / C] TGAAGCSAGGGATATCCTCATTCATTCTTTTGGTCTCAATTGCCCAGCATAATGCCAGGT 9 ALGA0056924 ATGGCGTGGTAAGGATGTGCATCAGGAGGCCAGCTGTCCTTCTCCTTACGTCTCTGTACC [A / G] ACCTTCCTACTTCGTCCAAAGCATTCATCCAGCGTTGCAAGGGCAATTGCTCAGACTCCA 10 ALGA0064322 AAGCTGAGGAAGAGCTCCCACGCTGGCCTCCTGTGGATGTCGCACCAGCTCGCGGGGCCC [T / C] CATGCCAGGACCTACCTCTAACTCCTCCTGGGTGGACACTCTGGTGGTCAGCAGAAGGAA 11 ALGA0064392 GCTGCAACCAGAGTCACAGAAGTGACAACACTGGTTCCTTATCCCGCTGAGCCATGAGAA [T / G] CTGTACTTTTAGAAGAGCTGAGTCGTCTCTTAGCTGCCTTAAACCTAGTCCTGGTCCCAA 12 ALGA0065426 CTGCCCTCCCCACTCCCTTCCTGGGCAGGTGAAGGGCAGGAAGTTAGCCTGGTCTAGCTT [T / C] CTCAGGTTAAACAGAAAGCTACAGGCTCACAGTCACCAGTACCTAGTAAAGTGGGGCAAG 13 ALGA0067483 ATGTTTTCTGAACCAAAGTAAGAACATGGGATCTGACAAATGTACTTATGGGGGACCTGG [A / G] GTGTGCTGTCCTTGAGACTGGAATTGGGACTCTTAGCCAAAATCACACAAAAATTAAGTA 14 ALGA0072858 AGCTTCCCTCCTAAAATTATTATCTATTAATAGTAATAATGATAATAGCTAACACATGTA [A / G] TGCTTACTATATGATAGGCACTATCCTAAGAACTCATGAAATGATTCAATATTCTTAGTT 15 ALGA0079359 TCAGGGAACAGAGGATTGGGGAGCCTGCAGTTTCTCTTCTGCCTCTCTGTGGCTCTTCCT [T / C] GCAGCTCAGGTCCAGGGATGATCTGGGACGGACTTTAGTGGCTATCTAGTCCAACTCTCT 16 ALGA0085130 CTCCTCTTTTGAACTGTGAGAAATAAACTTGTCTTTTCAGTTAAGATCATCTCCTGATCT [A / G] TCAGCTTTCCTCTACCTCAAAATTTTTATTAGTACTCTGTATTTTAAGACAAGATGTTGT 17 ALGA0089251 TACGGTTTACTTATCAGTGAGATGTCCCCCCTTAGTTTAGAGAGTTTCCTTACATATCTG [T / G] TATAATATCTGGCCCCAGAAGCTATGTCTTGGGGATAATATATATATATGATCCTCTGAT 18 ALGA0092844 CACCATTAGAGGGGATGCTCTCTAAGGCTGCTTCGTTCAAACTTCCTGATTCCAGCTGCA [T / C] GCAGGTAGGGAACAGAGCTGACCATGACCGAAGGACACTTGGAAATCCACATATTAGCAC 19 ALGA0097474 AGATGCCAAGATTACTTATCTCTGTCAAACTAGCTGCCCCAAGATGGCAGCTGGAAGATG [T / C] TTATGCCCTGCCCGTGGCAAGGTGAGCCACGGCAGGAATTACAGCTTGTGCTTGAAAATA 20 ALGA0097857 GGGTTGGCGAAAATCAAGGGTATCCTCAGAGAAGAGACTCAGACACAGCCCTCAAAGTGC [A / G] CACTCAGGCACTTGACGTAGCTTTAGTGCTACCTCTTCAATATCTTCGAGAGTGAGGAAA 21 ALGA0103611 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCTTCCAT [A / G] GGCAGCAAGTGACAATTCCGAATCATTATAACAACAGCAAAGTCAACGCCTTACACTTGA 22 ALGA0106326 GGCGCTTTGTGAAAGAGCCCCGTGTTGGCCCTTAAGGTGGCTTTGGACCTGCCTCTAGGT [A / G] ACCCTGAACCAGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 23 ALGA0110333 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCTGTTTAGTCTGTGTGG [T / C] TGAGGCCAAAAGCTTGTGCTGCCTCCGAGGGGAGCATCGCATAATTGCAGGCAGAGGGGG 24 ALGA0114065 TCTGGAGATTCTAGGAGAATAAGAGCATCCTACTGCTTTTCTATAGAATATGGGGATGCC [T / C] ATCACCTATCATGTCCTAGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 25 ALGA0114861 GTTGTTTCTGCATAAATGAGAAAATTGTGGCCAACTACAAAGTTTACATGTGTAATTTAG [T / G] TCTAATACTGGAAAAGATAGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 26 ALGA0123954 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCTGCCGTCCAGGA [T / C] GGGTCTCAGCCTCTTACCGGCCCTCCTCTCAAGCTTCCTGGTAACTTTGTGCGAACTCGA 27 ALGA0124374 CGTGCTGAGGGTCAGGAGGTGCAGGTGCATGACCCCCTCCCTTTTAACTTTGGTGCCCAG [A / G] CCTGTCCCTGCTCCTCCCAGGCCCAAGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 28 ASGA0040082 CCAGGAGGGAGACGTTCAAATAGGTTCCTCCAGCTACTTGCAAGTGAACTTGAAAAATGA [T / C] GGGCCACCCGCAGGACTCCTTTCTAACGTGACAGAGGCGGATGCATTTGCGCATCAGCGAG 29 ASGA0089719 TACTGACAACAAGCCGTGAACACGAATAGAAAAGACTTGGAAACAGTTCATACTGTGTGA [T / C] CGCACTGGCTGAGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN 30 ASGA0092931 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCTGCTGTAC [A / G] CACCCAGGGTTGACTTATTTTTTTTAGGTAGACACAGTGTCTGGGCCCATGAGAATGTTT 31 ASGA0096881 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCTGGGAGTGACAGGACATCC [T / C] GTCTCTCTCCTGATTCTAATCTGAGAGAGTCCTACCTCCAAGGCCCTTGGTACCTGCTGC 32 ASGA0102105 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCTCCGTGACCTG [A / G] TGGTAGCTGCCGAACCTTGCTGTCTCGGTTCCCTCTCTGTAGGACAGGGACAATGACCAC 33 H3GA0000077 GGTCCTCATGAGAACCCTAGGAGTAGATCGTTTAAAAGACTGTCCTTCATTCTGCAGAAG [A / C] GGCGGCCGAGCCCTGAGAGGTTACGTGAGTTTCCCCAGCTGCACAGTGGGCCCCTGACAG 34 H3GA0028278 GTTTTTCTGTTCTTTGATTTCTTAGATCAGGATCAGCAGACATTTTCCATCAAGGGCCAC [A / G] TAATAAATATTTTAGGCATGCAGTCTCTTTCACAGTTGCTCAGCTCTGCTTTTGTAGCCA 35 H3GA0031292 TTCATTCCTGGTTTGGGCAAAGACGTTGAATTGGCAGTGGGATTCCCTGTGATACTGGGT [A / G] AGGAACTCAGGGAATGGAGTGGGCCGAAGTTGAAGAAAATTGACTGTTTCCGCTTAGAAA 36 M1GA0000139 AAGCGGACGCCCACAGCCAGCGCGGAGCGCCCCAGGGTTCCGGGAAGCCGGTTTCTCCTC [T / C] GTGATGAGACACGGCAGCTTCAGACTCGGACGGAGTTTCACGGAACTTACATGCGGTGGA 37 M1GA0001903 GTTTTGCTATTTCCTGGGCTAAGACGGGGAGGCAGAGGGTCGAACCGGGAGCAGGGSATC [A / C] CTTCCCTTTTATTAAGTTTACGGTACCTGGGAAACACCTGAAGTGGAGAGACGCTAACGG 38 MARC0004720 AAAATTGCACAAAGGACCATAGGGGAAACCCAAAGGCTGGTTAGATAGGGTCTATTTTCT [T / C] CTTAGCCTATGAGTCATTTGGGGAAAAATTACAGAGGGCATAGTGATCTTTGATTTACCA

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Attach an electronic file to a sequence list

Claims (4)

A pig individual identification method comprising identifying a monobasic polymorphism, which is the 61st base sequence of the polynucleotides of SEQ ID NOs: 1 to 38.

The method of claim 1, wherein the step of identifying a single nucleotide polymorphism comprises allele-specific probe hybridization, allele-specific amplification, sequencing, and 5. 5 'nuclease digestion, molecular beacon assay, oligonucleotide ligation assay, size analysis and single-stranded polymorphism Individual identification method of pigs, characterized in that performed by a method selected from the group consisting of stranded conformation polymorphism.
A microarray for detecting a single nucleotide polymorphism comprising a polynucleotide for recognizing the nucleotide sequence of the single nucleotide polymorphism of claim 1, a polypeptide encoded therein, or a cDNA of the polynucleotide.
The kit for detecting a single nucleotide polymorphism comprising a polynucleotide for recognizing the nucleotide sequence of the single nucleotide polymorphism of claim 1, a polypeptide encoded therein or a cDNA of the polynucleotide.