CN105349626A - Identification method of Laiwu pig and its meat products - Google Patents
Identification method of Laiwu pig and its meat products Download PDFInfo
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Abstract
The invention belongs to a variety identification technique of livestock and their meat products and particularly relates to an identification method of Laiwu pig and its meat products. A D-loop area, Cytb gene and mtDNA total sequences of a pig to be identified can be amplified using primer sequences provided by the invention. A sequence library of the D-loop area and Cytb gene of all-population purebred Laiwu pigs has been established, and a GENBANK database of purebred Laiwu pigs in NCBI also has mtDNA total sequences at present. According to the principle that the pig to be identified and the Laiwu pig are different in mtDNA nucleotide, whether the pig to be identified and its meat products are from Laiwu pig or not can be judged accurately. The method can identify whether a pig variety or meat products are from Laiwu pig or not and can also judge approximately its variety source if the pig variety or meat products are not from Laiwu pig.
Description
Technical field
The invention belongs to livestock-raising technology, specifically the authentication method of a kind of Laiwu Pigs and meat product thereof.
Background technology
Laiwu Pigs is one of excellent local variety pig of China, and famous with higher intramuscular fat content (about 10%), it is the pig variety that a nutritive value and economic worth are all very high.Laiwu Pigs enjoys liking of culturist and human consumer because kind is unique, cause the counterfeit of a lot of Laiwu Pigs or its meat product on market.Accurately and efficiently identify that Laiwu Pigs or its meat product become a kind of means of urgent need, the authentication method of science can not only be provided, and can the boar of specification Laiwu Pigs and meat product market, and then safeguard the rights and interests of culturist and human consumer.
The method of pig variety qualification mainly contains judging of exterior method and Protocols in Molecular Biology method, and judging of exterior method needs experienced especially middle man, and mainly by seeing and touch etc. that subjective experience is evaluated pig kind appearance, have certain subjectivity, authentication method is science not.The kind methodological science of Protocols in Molecular Biology qualification pig, reliable results, but need the specific molecular marker finding this kind applicable.
The kind of molecular biology identification pig mainly contains three kinds of methods: one is use the mtDNA of matrilinear inheritance as mark, and this is also the most frequently used method, because the rich polymorphism of each pig variety mtDNA, gene fragment is shorter.Two is patroclinous Y chromosomes, and this method uses less, because the top grade species specific marker site of pig Y chromosome is little.Three is select the microsatellite marker on nuclear chromosome, and this method is relatively reliable, but the time that the species specific microsatellite marker of the product that are to locate needs is long, and the cost expended is high.Up to the present, be the most frequently used method with mtDNA as molecular markers for identification kind, but because of its rich polymorphism, so will set up all polymorphic site storehouses of this kind mtDNA, the research about Laiwu Pigs kind and meat product qualification thereof is very few.
Summary of the invention
The object of the present invention is to provide the authentication method of a kind of Laiwu Pigs and meat product thereof.
For achieving the above object, the technical solution used in the present invention is:
An authentication method for Laiwu Pigs and meat product thereof, by pig to be identified or the sampling of its meat product, the primer provided by present method carries out amplification and checks order, and can identify that whether this pig kind is whether Laiwu Pigs or its meat product are from Laiwu Pigs by sequence alignment.
Pig to be identified or its meat product are from purebred pig, if hybridized pig, the method can only identify whether its mother derives from Laiwu Pigs.
Extract STb gene after described pig to be identified or the sampling of its meat product, first the primer provided by present method carries out the amplification order-checking of D-loop district and Cytb gene, and full group's gene pool of the Laiwu Pigs D-loop district provided with present method and Cytb gene is compared.
According to the comparison result of D-loop district and Cytb gene, described tentatively judges that whether pig to be identified is from Laiwu Pigs, if "no", jumps at a conclusion, if "Yes", further verify with the mtDNA total order primer that present method provides further.
The described mtDNA gene total order obtaining pig to be identified with the mtDNA total order primer that present method provides, then with the sequence alignment in GENBANK gene pool, verify that whether pig to be identified or its meat product are from Laiwu Pigs.
With mtDNA total order primer, described accurately can judge that whether this purebred pig or its meat product are from Laiwu Pigs, if not from Laiwu Pigs, can roughly judge its cultivar origin by the comparison of mtDNA total order.
The advantage that the present invention has:
Only amplification D-loop district and Cytb gene check order, and cost is lower, process provides the polymorphic site storehouse of Laiwu Pigs D-loop district and Cytb gene, can get rid of pig to be identified very soon and meat product is not from Laiwu Pigs.The D-loop district that present method provides, Cytb gene and mtDNA total order primer are the universal primers of pig, as long as the sample of pig can increase out.
Accompanying drawing explanation
Fig. 1 pig D-loop to be identified district and Cytb gene and Laiwu Pigs gene pool comparison result, can judge that this pig or meat product are not Laiwu Pigs.This qualification pig D-loop district (figure A) and Cytb gene (figure B) all do not have identical sequence with the comparison of Laiwu Pigs gene pool.
Fig. 2 pig D-loop to be identified district and Cytb gene and Laiwu Pigs gene pool comparison result, can this pig of preliminary judgement or meat product from Laiwu Pigs, need may verify by mtDNA total order further.(A figure) is identical with Laiwu Pigs D-loop3 sequence in this qualification pig D-loop district, and Cytb gene (B figure) is identical with Laiwu Pigs Cytb2 sequence.
The result figure of Fig. 3 Laiwu Pigs mtDNA provided by the invention total order and the comparison of GENBANK database, visible Laiwu Pigs has special mtDNA marker site.
Embodiment
In the following examples, the present invention is further elaborated, but the present invention is not limited thereto.
The intramuscular fat content of Laiwu Pigs is high, and meat is fragrant and delicious, causes Laiwu Pigs that market is much palmed off or its meat product.MtDNA has the feature of matrilinear inheritance, and rich polymorphism, testing cost is low, can be advantageously applied in animal varieties qualification.
Embodiment
In the present embodiment, the main purpose of Laiwu Pigs and meat product qualification thereof judges that whether pig to be identified or its meat product are from Laiwu Pigs, for the person of raising pigs or human consumer provide the foundation of science.Laiwu Pigs in the present invention and meat product authentication method thereof comprise the following steps:
1) sample
Take hair root or meat product (preferably raw meat) 3-5g of the ear tissue of pig to be identified, blood, band hair follicle, if can not carry out immediately testing, should sample is immersed in dehydrated alcohol, blood will add antithrombotics and 4 ° of C preserve.
2) extract DNA and carry out pcr amplification order-checking
Institute's sample thief phenol and chloroform method extracts STb gene, the D-loop district primer 1 provided by present method, F:5'-AGGAGACTAACTCCGCCAT-3', R:5'-CTGAGTCCAAGCATCCCCAA-3', and primer 2, F:5'-CGCGCATATAAGCAGGTAAA-3', R:5'-CGCGGATACTTGCATGTGT-3'; The primers F of Cytb gene: 5'-GGAATTTAACCACGACCAATGAC-3', R:5'-GTGGCCCTCCTTTTCTGGTTTAC-3'; MtDNA total order primer is in table 1.Conveniently PCR system increases, annealing temperature 58-64 ° of C, checks order after amplification with PCR liquid.
Table 1 increases the primer information of mtDNA total order
| Primer numbers | Primer sequence 5'-3'(upstream/downstream) |
| 1 | AGGAGACTAACTCCGCCAT/ CGCGGATACTTGCATGTGT |
| 2 | ACTAAGTCAATGCCTATTCTG/CAAATGTATGAAACCTCAG |
| 3 | CTACACAATAACCTCCCATA/TGGCACGAGATTTACCAACT |
| 4 | GCTCATAACGCCTTGCTC/ATTCTTTCATCTTTCCCTT |
| 5 | CACCTAGAAGATCCCACA/ACAACCAGCTATCACCAG |
| 6 | CCGTAAGGGAAAGATGAAAG/TATGGTTATTTTGACTGGT |
| 7 | CCGTGCAAAGGTAGCATA/CCAACATCGAGGTCGTAA |
| 8 | TGGGGTGACCTCGGAGTAC/AATATGGCGAAAGGTCCGG |
| 9 | CGAGCAGTAGCCCAAACA/GGTCGTATCGGAATCGTG |
| 10 | GTATCAGGCTTTAACGTAGA/TGGTAATACTGCTGTCATTC |
| 11 | CACAGAAGCAGCCACAAA/ATGGGATAGGGATAAAGT |
| 12 | ACATAGGATGAATGACAGC/TGGTGGAAGTAGTCAGAAAC |
| 13 | GCACTGCCTTGAGCCTAC/GTGTTCAGGTTGCGGTCT |
| 14 | CTGACTCGTACCGCTAATA/CTGACTCGTACCGCTAATA |
| 15 | CACTTTGTAATCATATTCGTAG/TAGTTGGAAAGGGTAAGC |
| 16 | TTCATCTCACTAACAGCAG/TTGAGTTCGGTTGATTCTG |
| 17 | GCTTCATGCCCATTGTAC/TTATAGCGGAATCCTGTG |
| 18 | GCAAGCCCAGAATCAACCG/CGAGGAGGATTGAGGTGTT |
| 19 | ATACCACATAGTAAACCCAA/CCTGTAGCCACAAAGAAA |
| 20 | CTAAACACCTCAATCCTCC/TTGGACGTAATCGGTACCG |
| 21 | CCTTGCAGGGTTACTTAT/TTCGGGTTGTGGTTTCTT |
| 22 | CGGTACCGATTACGTCCAA/CCGATTAGATTGATGGATG |
| 23 | ACCAGCTCTATCTGCTTA/GAGGCTTTGATGTTGTTA |
| 24 | ATGATGACTAATAGCAAGCC/GGGATGTAGTCCGAATTG |
| 25 | CATCGGAGACATTGGATT/AGTTGGCTTGAAGTTGAG |
| 26 | CCTACTCCTAGCTGCAGCAG/ATTATGGAGATTACTCGTGG |
| 27 | TCCGCATCATCATTACTA/TTTATGGTGGACTTGGGT |
| 28 | TAATTACCACGAGTAATCTC/TTCTACGAGGTCTGTTCCG |
| 29 | GGAGCATCCATATTCTTT/GGTGTAGTTGTCTGGGTCT |
| 30 | TCGTAGAATGAATCTGAGG/GGTGATACGCATGTTGACTG |
3) whether D-loop district and Cytb gene order identify be from Laiwu Pigs
The Laiwu Pigs sequence library DNAMAN software that the D-loop district of the pig to be identified obtained and Cytb gene provide with present method is respectively carried out sequence alignment, if there is no the sequence (see figure 1) of the type in the sequence library provided, then can judge that pig to be identified is not from Laiwu Pigs; If there is the sequence (see figure 2) of the type in the sequence library provided, then use the mtDNA total order of mtDNA total order primer amplification pig to be identified.
4) whether the qualification of mtDNA total order is from Laiwu Pigs
After obtaining the mtDNA total order of pig to be identified, comparison in GENBANK database, concrete steps are: NCBI-BLAST-nucleotideblast-pig to be identified mtDNA total order-database selects Nucleotidecollection (nr/nt)-source of species (Susscrofa (taxid:9823))-Optimizefor (Somewhatsimilarsequences (blastn))-FiltersandMasking not establish any parameter, and other parameters are default value.Then can judge that whether pig to be identified is from Laiwu Pigs, also can roughly judge its cultivar origin according to sequence homology size.Comparison result is shown in Fig. 3.
LaiwuD-loop1
CAACCAAAACAAGCATTCCATTCGTATGCAAACCAAAACGCCAAGTACTTAATTACTATCTTTAAAACAAAAAAACCCATAAAAATTGCGCACAAACATACAAATATGCGACCCCAAAAATTTAACCATTAAAAACAAAAAATTTAATATATTATAGCCCTATGTACGTCGTGCATTAACTGCTAGTCCCCATGCATATAAGCATGTACATATTATTATTAATATTACATAGTACATATCATTATTGATCGTACATAGCACATATCATGTCAAATAATTCCAGTCAACATGCGTATCACCACCATTAGATCACGAGCTTAATTACCATGCCGCGTGAAACCAGCAACCCGCTTGGCAGGGATCCCTCTTCTCGCTCCGGGCCCATGAATCGTGGGGGTTTCTATTGATGAACTTTAACAGGCATCTGGTTCTTACTTCAGGACCATCTCACCTAAAATCGCCCACTCTTTCCCCTTAAATAAGACATCTCGATGGACTAATGACTAATCAGCCCATGCTCACACATAACTGAGGTTTCATACATTTGGTATTTTTTAATTTTTGGGGATGCTTGGACTCAGCCATGGCCGTCAAAGGCCCTAACACAGTCAAATCAATTGTAGCTGGACTTCATGGAACTCATGATCCGGCACGACAATCCAAACAAGGTGCTATTCAGTCAATGGTTACGGGACATAACGTGCGTACACGCGCATATAAGCAGGTAAATTATTAGCTCATTCAAACCCCCCTTACCCCCCATTAAACTTATGCTCTACACACCCTATAACGCCTTGCCAAACCCCAAAAACAAAGCAGAGTGTACAAATACAATAAGCCTAACTTACACTAAACAACATTTAACAACACAAACCACCATATCTTATAAAACACTTACTTAAATACGTGCTACGAAAGCAGGCACCTACCCCCCTAGATTTTTACGCCAATCTACCACAAATAAGTTTAAAATTACAACACAATAACCTCCCAAAATATAAGCACCTATTTAAGCATACGCCCACAATCTGAATATAGCTTATA
LaiwuD-loop2
CAACCAAAACAAGCATTCCATTCGTATGCAAACCAAAACGCCAAGTACTTAATTACTATCTTTAAAACAAAAAAACCCATAAAAATTGCGCACAAACATACAAATATGCGACCCCAAAAATTTAACCATTAAAAACAAAAAATTTAATATATTATAGCCCTATGTACGTCGTGCATTAACTGCTAGTCCCCATGCATATAAGCATGTACATATTATTATTAATATTACATAGTACATATCATTATTGATCGTACATAGCACATATCATGTCAAATAACTCCAGTCAACATGCGTATCACCACCATTAGATCACGAGCTTAATTACCATGCCGCGTGAAACCAGCAACCCGCTTGGCAGGGATCCCTCTTCTCGCTCCGGGCCCATAAATCGTGGGGGTTTCTATTGATGAACTTTAACAGGCATCTGGTTCTTACTTCAGGACCATCTCATCTAAAATCGCCCACTCTTTCCCCTTAAATAAGACATCTCGATGGACTAATGACTAATCAGCCCATGCTCACACATAACTGAGGTTTCATACATTTGGTATTTTTTAATTTTTGGGGATGCTTGGACTCAGCCATGGCCGTCAAAGGCCCTAACACAGTCAAATCAATTGTAGCTGGACTTCATGGAACTCATGATCCGGCACGACAATCCAAACAAGGTGCTATTCAGTCAATGGTTACGGGACATAACGTACGTACACGCGCATATAAGCAGGTAAATTATTAGCTCATTCAAACCCCCCTTACCCCCCATTAAACTTATGCTCTACACACCCTATAACGCCTTGCCAAACCCCAAAAACAAAGCAGAGTGTACAAATACAATAAGCCTAACTTACACTAAACAACATTTAACAACACAAACCACCATATCTTATAAAACACTTACTTAAATACGTGCTACGAAAGCAGGCACCTACCCCCCTAGATTTTTACGCCAATCTACCACAAATAAGTTTAAAATTACAACACAATAACCTCCCAAAATATAAGCACCTATTTAAGCATACGCCCACAATCTGAATATAGCTTATA
LaiwuD-loop3
CAACCAAAACAAGCATTCCATTCGTATGCAAACCAAAACGCCAAGTACTTAATTACTATCTTTAAAACAAAAAAACCCATAAAAATTGCGCACAAACATACAAATATGCGACCCCAAAAATTTAACCATTAAAAACAAAAAATTTAATATATTATAGCCCTATGTACGTCGTGCATTAACTGCTAGTCCCCATGCATATAAGCATGTACATATTATTATTAATATTACATAGTACATATTATTATTGATCGTACATAGCACATATCATGTCAAATAATTCCAGTCAACATGCGTATCACCACCATTAGATCACGAGCTTAATTACCATGCCGCGTGAAACCAGCAACCCGCTTGGCAGGGATCCCTCTTCTCGCTCCGGGCCCATAAATCGTGGGGGTTTCTATTGATGAACTTTAACAGGCATCTGGTTCTTACTTCAGGACCATCTCACCTAAAATCGCCCACTCTTTCCCCTTAAATAAGACATCTCGATGGACTAATGACTAATCAGCCCATGCTCACACATAACTGAGGTTTCATACATTTGGTATTTTTTAATTTTTGGGGATGCTTGGACTCAGCCATGGCCGTCAAAGGCCCTAACACAGTCAAATCAATTGTAGCTGGACTTCATGGAACTCATGATCCGGCACGACAATCCAAACAAGGTGCTATTCAGTCAATGGTTACGGGACATAACGTACGTACACGCGCATATAAGCAGGTAAATTATTAGCTCATTCAAACCCCCCTTACCCCCCATTAAACTTATGCTCTACACACCCTATAACGCCTTGCCAAACCCCAAAAACAAAGCAGAGTGTACAAATACAATAAGCCTAACTTACACTAAACAACATTTAACAACACAAACCACCATATCTTATAAAACACTTACTTAAATACGTGCTACGAAAGCAGGCACCTACCCCCCTAGATTTTTACGCCAATCTACCACAAATAAGTTTAAAATTACAACACAATAACCTCCCAAAATATAAGCACCTATTTAAGCATACGCCCACAATCTGAATATAGCTTATA
LaiwuD-loop4
CAACCAAAACAAGCATTCCATTCGTATGCAAACCAAAACGCCAAGTACTTAATTACTATCTTTAAAACAAAAAAACCCATAAAAATTGCGCACAAACATACAAATATGCGACCCCAAAAATTTAACCATTAAAAACAAAAAATTTAATATATTATAGCCCTATGTACGTCGTGCATTAACTGCTAGTCCCCATGCATATAAGCATGTACATATTATTATTAATATTACATAGTACATATCATTATTGATCGTACATAGCACATATCATGTCAAATAATTCCAGTCAACATGCGTATCACCACCATTAGATCACGAGCTTAATTACCATGCCGCGTGAAACCAGCAACCCGCTTGGCAGGGATCCCTCTTCTCGCTCCGGGCCCATAAATCGTGGGGGTTTCTATTGATGAACTTTAACAGGCATCTGGTTCTTACTTCAGGGCCATCTCACCTAAAATCGCCCACTCTTTCCCCTTAAATAAGACATCTCGATGGACTAATGACTAATCAGCCCATGCTCACACATAACTGAGGTTTCATACATTTGGTATTTTTTAACTTTTGGGGATGCTTGGACTCAGCCATGGCCGTCAAAGGCCCTAACACAGTCAAATCAATTGTAGCTGGACTTCATGGAACTCATGATCCGGCACGACAATCCAAACAAGGTGCTATTCAGTCAATGGTTACAGGACATAACGTGCGTACACGCGCATATAAGCAGGTAAATTATTAGCTCATTCAAACCCCCCTTACCCCCCATTAAACTTATGCTCTACACACCCTATAACGCCTTGCCAAACCCCAAAAACAAAGCAGAGTGTACAAATACAATAAGCCTAACTTACACTAAACAACATTTAACAACACAAACCACCATATCTTATAAAACACTTACTTAAATACGTGCTACGAAAGCAGGCACCTACCCCCCTAGATTTTTACGCCAATCTACCACAAATAAGTTTAAAATTACAACACAATAACCTCCCAAAATATAAGCACCTATTTAAGCATACGCCCACAATCTGAATATAGCTTATA
LaiwuCytb1
ATGACCAACATCCGAAAATCACACCCACTAATAAAAATTATCAACAACGCATTCATTGACCTCCCAGCCCCCTCAAACATCTCATCATGATGAAACTTCGGTTCCCTCTTAGGCATCTGCCTAATCTTGCAAATCCTAACAGGCCTGTTCTTAGCAATACATTACACATCAGACACAACAACAGCTTTCTCATCAGTTACACACATCTGTCGAGACGTAAATTACGGATGAGTTATTCGCTACCTACATGCAAACGGAGCATCCATGTTCTTTATTTGCCTATTCATCCACGTAGGCCGAGGCCTATACTACGGATCCTATATATTCCTAGAAACATGAAACATTGGAGTAGTCCTACTATTTACCGTTATAGCAACAGCCTTCATAGGCTACGTCCTGCCCTGAGGACAAATATCATTCTGAGGAGCTACGGTCATCACAAATCTACTATCAGCTATCCCTTATATCGGAACAGACCTCGTAGAATGAATCTGAGGGGGCTTTTCCGTCGACAAAGCAACCCTCACACGATTCTTCGCCTTTCACTTTATCCTGCCATTCATCATTACCGCCCTCGCAGCCGTACATCTCCTATTCCTGCACGAAACCGGATCCAACAACCCTACCGGAATCTCATCAGACATAGACAAAATTCCATTTCACCCATACTACACTATTAAAGACATTCTAGGGGCCTTATTTATAATACTAATCCTACTAATCCTTGTACTATTCTCACCAGACCTACTAGGAGACCCAGACAACTACACCCCAGCAAACCCACTAAACACCCCACCCCATATTAAACCAGAATGATATTTCTTATTCGCCTACGCTATCCTACGTTCAATTCCTAATAAACTAGGTGGAGTGCTAGCTCTAATAGCCTCCATCCTAATCCTAATTTTAATGCCCATACTACACACATCCAAACAACGAAGCATAATATTTCGACCACTAAGTCAATGCCTATTCTGAATACTAGTAGCAGACCTCATTACACTAACATGAATTGGAGGACAACCCGTAGAACACCCATTCATCATCATCGGCCAACTAGCCTCCATCTTATATTTCCTAATCATTCTAGTATTGATACCAATCACTAGCATCATCGAAAACAACCTATTAAAGTGAAGA
LaiwuCytb2
ATGACCAACATCCGAAAATCACACCCACTAATAAAAATTATCAACAACGCATTCATTGACCTCCCAGCCCCCTCAAACATCTCATCATGATGAAACTTCGGTTCCCTCTTAGGCATCTGCCTAATCTTGCAAATCCTAACAGGCCTGTTCTTAGCAATACATTACACATCAGACACAACAACAGCTTTCTCATCAGTTACACACATCTGTCGAGACGTAAATTACGGATGAGTTATTCGCTACCTACATGCAAACGGAGCATCCATGTTCTTTATTTGCCTATTCATCCACGTAGGCCGAGGCCTATACTACGGATCCTATATATTCCTAGAAACATGAAACATTGGAGTAGTCCTACTATTTACCGTTATAGCAACAGCCTTCATAGGCTACGTCCTGCCCTGAGGACAAATATCATTCTGAGGAGCTACGGTCATCACAAATCTACTATCAGCTATCCCTTATATCGGAACAGACCTCGTAGAATGAATCTGAGGGGGCTTTTCCGTCGACAAAGCAACCCTCACACGATTCTTCGCCTTTCACTTTATCCTGCCATTCATCATTACCGCCCTCGCAGCCGTACATCTCCTATTCCTGCACGAAACCGGATCCAACAACCCTACCGGAATCTCATCAGACATAGACAAAATTCCATTTCACCCATACTACACTATTAAAGACATTCTAGGGGCCTTATTTATAATACTAATCCTACTAATCCTTGTACTATTCTCACCAGACCTACTAGGAGACCCAGACAACTACACCCCAGCAAACCCACTAAACACCCCACCCCATATTAAACCAGAATGATATTTCTTATTCGCCTACGCTATCCTACGTTCAATTCCTAATAAACTAGGTGGAGTGCTAGCTCTAATAGCCTCCATCCTAATCCTAATTTTAATGCCCATACTACACACATCCAAACAACGAAGCATAATATTTCGACCACTAAGTCAATGCCTATTCTGAATACTAGTAGCAGACCTCATTACACTAACATGAATTGGAGGACAACCCGTAGAACACCCATTCATCGTCATCGGCCAACTAGCCTCCATCTTATATTTCCTAATCATTCTAGTATTGATACCAATCACTAGCATCATCGAAAACAACCTATTAAAGTGAAGA
LaiwuCytb3
ATGACCAACATCCGAAAATCACACCCACTAATAAAAATTATCAACAACGCATTCATTGACCTCCCAGCCCCCTCAAACATCTCATCATGATGAAACTTCGGTTCCCTCTTAGGCATCTGCCTAATCTTGCAAATCCTAACAGGCCTGTTCTTAGCAATACATTACACATCAGACACAACAACAGCTTTCTCATCAGTTACACACATCTGTCGAGACGTAAATTACGGATGAGTTATTCGCTACCTACATGCAAACGGAGCATCCATGTTCTTTATTTGCCTATTCATCCACGTAGGCCGAGGCCTATACTACGGATCCTATATATTCCTAGAAACATGAAACATTGGAGTAGTCCTACTATTTACCGTTATAGCAACAGCCTTCATAGGCTACGTCCTGCCCTGAGGACAAATATCATTCTGAGGAGCTACGGTCATCACAAATCTACTATCAGCTATCCCTTATATCGGAACAGACCTCGTAGAATGAATCTGAGGGGGCTTTTCCGTCGACAAAGCAACCCTCACACGATTCTTCGCCTTTCACTTTATCCTGCCATTCATCATTACCACCCTCGCAGCCGTACATCTCCTATTCCTGCACGAAACCGGATCCAACAACCCTACCGGAATCTCATCAGACATAGACAAAATTCCATTTCACCCATACTACACTATTAAAGACATTCTAGGGGCCTTATTTATAATACTAATCCTACTAATCCTTGTACTATTCTCACCAGACCTACTAGGAGACCCAGACAACTACACCCCAGCAAACCCACTAAACACCCCACCCCATATTAAACCAGAATGATATTTCTTATTCGCCTACGCTATCCTACGTTCAATTCCTAATAAACTAGGTGGAGTGCTAGCTCTAATAGCCTCCATCCTAATCCTAATTTTAATGCCCATACTACACACATCCAAACAACGAAGCATAATATTTCGACCACTAAGTCAATGCCTATTCTGAATACTAGTAGCAGACCTCATTACACTAACATGAATTGGAGGACAACCCGTAGAACACCCATTCATCATCATCGGCCAACTAGCCTCCATCTTATATTTCCTAATCATTCTAGTATTGATACCAATCACTAGCATCATCGAAAACAACCTATTAAAGTGAAGA
Claims (7)
1. an authentication method for Laiwu Pigs and meat product thereof, is characterized in that: by pig to be identified or the sampling of its meat product, the primer provided by present method carries out amplification order-checking, can identify that whether this pig or its meat product are from Laiwu Pigs by sequence alignment.
2. press the authentication method of Laiwu Pigs according to claim 1 and meat product thereof, it is characterized in that: pig to be identified or its meat product are from purebred pig, if hybridized pig, the method can only identify whether its mother derives from Laiwu Pigs.
3. by the authentication method of Laiwu Pigs described in claim 1 or 2 and meat product thereof, it is characterized in that: after pig to be identified or the sampling of its meat product, extract STb gene, first the primer provided by present method carries out the amplification order-checking of D-loop district and Cytb gene, and full group's gene pool of the Laiwu Pigs D-loop district provided with present method and Cytb gene is compared.
4. press the authentication method of Laiwu Pigs according to claim 3 and meat product thereof, it is characterized in that: the comparison result according to D-loop district and Cytb gene tentatively judges that whether pig to be identified is from Laiwu Pigs, if "no", jump at a conclusion, if "Yes", further verify with the mtDNA total order primer that present method provides further.
5. according to the authentication method of the Laiwu Pigs described in right 4 and meat product thereof, it is characterized in that: the mtDNA total order primer provided by present method obtains the mtDNA gene total order of pig to be identified, again with the sequence alignment in GENBANK gene pool, verify that whether pig to be identified or its meat product are from Laiwu Pigs.
6. the Laiwu Pigs according to right 1 and 5 and meat product authentication method thereof, it is characterized in that: accurately can judge that whether this purebred pig or its meat product are from Laiwu Pigs with mtDNA total order primer, if not from Laiwu Pigs, can roughly judge its cultivar origin by the comparison of mtDNA total order.
7. the Laiwu Pigs according to right 3 and the authentication method of meat product thereof, it is characterized in that: only amplification D-loop district and Cytb gene check order, the full group's Laiwu Pigs D-loop district provided with present method and the storehouse comparison of Cytb gene order, cost is lower, if pig to be identified is not can conclude very soon from Laiwu Pigs.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012229A (en) * | 2017-04-24 | 2017-08-04 | 吉林农业科技学院 | Pig derived component quick determination method and kit in food |
| CN110331212A (en) * | 2019-07-09 | 2019-10-15 | 河南农业大学 | A method of the black pig in Queshan is identified using D-loop region mutation site |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101715488A (en) * | 2008-06-26 | 2010-05-26 | 马来西亚博特拉大学 | A method for identifying a pork content in a food |
| CN101864476A (en) * | 2009-04-15 | 2010-10-20 | 江苏省淡水水产研究所 | Method for identifying coilia ectenes and coilia mystus by using PCR |
| CN102206712A (en) * | 2011-04-27 | 2011-10-05 | 中国水产科学研究院东海水产研究所 | Method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence |
| CN103215365A (en) * | 2013-04-26 | 2013-07-24 | 广西大学 | DNA (Deoxyribonucleic Acid) molecular identification method for Luchuan pigs and meat products thereof |
-
2015
- 2015-10-09 CN CN201510647370.9A patent/CN105349626A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101715488A (en) * | 2008-06-26 | 2010-05-26 | 马来西亚博特拉大学 | A method for identifying a pork content in a food |
| CN101864476A (en) * | 2009-04-15 | 2010-10-20 | 江苏省淡水水产研究所 | Method for identifying coilia ectenes and coilia mystus by using PCR |
| CN102206712A (en) * | 2011-04-27 | 2011-10-05 | 中国水产科学研究院东海水产研究所 | Method for identifying pacific mackerel and slimy mackerel on basis of mitochondrial cytochrome b sequence |
| CN103215365A (en) * | 2013-04-26 | 2013-07-24 | 广西大学 | DNA (Deoxyribonucleic Acid) molecular identification method for Luchuan pigs and meat products thereof |
Non-Patent Citations (7)
| Title |
|---|
| GUANGHUI YU等: "The phylogenetic status of typical Chinese native pigs: analyzed by Asian and European pig mitochondrial genome sequences", 《JOURNAL OF ANIMAL SCIENCE AND BIOTECHNOLOGY》 * |
| HASSAN BARAKAT 等: "Detection of pork adulteration in processed meat by species-specific PCR-QIAxcel procedure based on D-loop and cytb genes", 《APPL MICROBIOL BIOTECHNOL》 * |
| JIYING WANG等: "Phylogenetic Relationships of Pig Breeds from Shandong Province of China and Their Influence by Modern Commercial Breeds by Analysis of Mitochondrial DNA Sequences", 《ITALIAN JOURNAL OF ANIMAL SCIENCE》 * |
| ZHANG，D.等: "Sus scrofa breed Lai wu mitochondrion，complete genome", 《GENBANK DATABASE》 * |
| 王继英等: "山东猪种mtDNA D-loop区部分序列遗传多样性及系统进化研究", 《畜牧兽医学报》 * |
| 王继英等: "山东省猪种mtDNA CytB基因遗传多样性及系统进化研究", 《中国农业科学》 * |
| 赵兴波 等: "猪线粒体DNA D-loop的遗传变异分析", 《遗传》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012229A (en) * | 2017-04-24 | 2017-08-04 | 吉林农业科技学院 | Pig derived component quick determination method and kit in food |
| CN110331212A (en) * | 2019-07-09 | 2019-10-15 | 河南农业大学 | A method of the black pig in Queshan is identified using D-loop region mutation site |
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