KR20170027078A - Composition for discrimination of Yeonsan Ogye - Google Patents

Abstract

The present invention relates to a composition for discriminating Yeonsan Ogye (all black chicken from Korea) using a native-specific gene, a kit, and a method for discriminating Yeonsan Ogye using the same. According to the present invention, the composition, the kit, and the method for discriminating breeds of chickens can discriminate Yeonsan Ogye, which is a rare breed of native chickens through a simple process, thereby being used for studying and preserving Yeonsan Ogye and discriminating a breed of Yeonsan Ogye among crossbreed chickens.

Description

[Composition for Discrimination of Yeonsan Ogye]

The present invention relates to a composition for identifying a gene coding sequence using a genetically-specific gene, a kit, and a method for identifying a gene coding sequence using the same.

Recognizing the value of conservation and utilization of native livestock as a genetic resource in different parts of the world (Notter (1999) J. Anim. Sci. 77: 61-69) And the relationship between them. For the breed of cattle, studies have been performed using various genetic markers such as blood proteins, milk proteins, microsatellites, mitochondrial DNA (MacHugh et al (1998) Anim. Genet. 29: Genetics 150: 1169-1175; Edwards et al (2000) Anim. Genet. 31: 127-130; Troy et al (2001) Nature 410: 1088-1091; Hanotte et al 2000) Molecular Ecology 9: 387-396, (2002) Science 296: 336-339).

Among these genetic markers, microsatellite is a repetitive DNA group consisting of about 2 to 6 nucleotides. It is distributed in the genome and has a highly non-coding DNA sequence, coding DNA sequence) (Zajc I and Sampson J (1999) J Hered 90: 104-107). Variability among individuals is recognized according to the number of repeats in a particular locus (Koreth J, O'Leary JJand McGee JO (1996) J Pathol 178: 239-248.), (Polymerase Chain Reaction (PCR)) using a primer designed in the region, it is possible to observe the polymorphism in the PCR product length and to detect the DNA polymorphism. In addition, microsatellite genes are transferred to their offspring according to Mendel's law of genetics, and can be amplified using PCR, and co-dominant when electrophoresed. In particular, since microsatellite is small in size, it can amplify two to eight primers simultaneously (Chamberlain JS et al., (1988) Nucleic Acids Res 16: 11141-11156) It saves time and money, and results in high sensitivity and accuracy. Thus, such microsatellite is very useful in the field of molecular breeding of livestock, and is useful not only to prevent degradation by inbreeding but also to select specific traits.

On the other hand, chickens are the representative livestock of Korea, and they are economically and industrially advantageous because they are faster in production cycle and relatively easy to transport than other livestock. According to the 2011 statistics, the consumption of chicken and eggs is rapidly increasing, with 11.4 kg of chicken per person and 232 eggs. However, compared with the growth of the poultry industry, the ratio of conventional chickens is only about 10% of the total domestic market.

Domestic native livestock genetic resources have been preserved and evaluated at livestock genetic resource test sites and management institutions. However, some resources that have been preserved for a long time in the farms have not been systematically managed by the state because they do not meet the related regulations. Hwanggeum, sage green, fowl, and Hwangbong are found in ancient literature and calligraphy, and farmers have been collecting it for more than 20 years and are trying to preserve the species. Regarding this, the Rural Development Administration added twenty-four varieties of five genera, including five genera, sage green, fowl, and hwangbong, to the Domestic Animal Diversity Information System of FAO in 2012.

Therefore, there is a need to develop a new breed of domestic native chicken as well as to genetically protect it. Korean Patent Laid-Open No. 10-2007-0051594 discloses a selective breeding using the mutation region of the deduced protein gene of Korean native chicken. It detects 10 base mutation regions in Korean traditional chicken UCP gene, However, the present invention does not disclose various genetic markers for distinguishing specific domestic varieties, such as the five genera.

In other words, there has not been much research on genetic markers that can genetically identify genetic markers effectively, and it is necessary to study for easy genetic identification and conservation of varieties using conventional chickens.

The inventors of the present invention discovered five kinds of genetic markers that specifically change the expression of the five genomic sequences while studying genetic markers capable of specifically discriminating native chicken cultivars and completed the present invention.

Accordingly, it is an object of the present invention to provide a composition for identifying a breed of chicken, a kit, and a method for discriminating a chicken breed using the same.

In order to achieve the above object, the present invention provides a biomarker comprising one or more genes selected from SEQ ID NOS: 4 to 8, a nucleic acid or oligonucleotide specific to the gene, or an antibody specific for a protein encoded by the gene , And a composition for distinguishing a breed of a chicken.

The present invention also provides a kit for identifying a breed of chicken comprising a biomarker consisting of one or more genes selected from SEQ ID NOS: 4 to 8, a nucleic acid or oligonucleotide specific to the gene, or an antibody specific to the protein encoded by the gene to provide.

The present invention also provides a microarray chip for discriminating a breed of chicken, in which at least one gene selected from SEQ ID NOS: 4 to 8 or a nucleic acid sequence complementary to the gene is integrated.

1) obtaining a gene sample from a chicken; 2) comparing the degree of expression of one or more genes selected from SEQ ID NO: 4 to SEQ ID NO: 8 in the sample of the first step; The present invention provides a method for distinguishing a chicken breed.

The composition, kit and method for discriminating breeds of chicken according to the present invention can discriminate the five kinds of native breeds, which are natural monuments and native breeds, through the simple process. Therefore, the breeding and conservation of the five breeding species, And the like.

FIG. 1 is a graph showing the result of confirming specific overexpression of CRLF2 (SEQ ID NO: 5) (WL: White Leghorn, YO: Computational Five, HIBL: Hypnotic, HB: Hwangbong, HY: Hoengseong).

The present invention provides a composition for identifying a breed of chicken, which comprises a biomarker consisting of one or more genes selected from SEQ ID NOS: 4 to 8, a nucleic acid or oligonucleotide specific to the gene, or an antibody specific to a protein encoded by the gene to provide.

In the present invention, it is preferable that the chicken is a fifth ovine.

The one or more genes selected from SEQ ID NOS: 4 to 8 of the present invention specifically change the expression in the five genes of the present invention, and thus can be distinguished from non-native species such as white legghorn and other native species such as Hoengseong chicken, Make it easy to identify the five lines.

The one or more genes selected from SEQ ID NOS: 4 to 8 may be used alone to discriminate a chicken breed, but when used in combination, the accuracy of chicken breed identification can be further improved.

The nucleic acid or the oligonucleotide specific to the gene may be a primer or a probe and may have a sequence complementary to a gene such as DNA and / or RNA, or hybridize.

The primer refers to a nucleic acid sequence capable of complementarily binding with a template and having a free 3 'hydroxyl group that allows the reverse transcriptase or DNA polymerase to initiate replication of the template . A primer is a nucleotide having a sequence complementary to the nucleotide sequence of a specific gene, and has a length of about 7 bp to 50 bp, preferably about 10 bp to 30 bp. The primers may also contain additional base sequences that do not alter the priming properties of the primers that serve as a starting point for DNA synthesis. The primers can be chemically synthesized using well-known methods, and such nucleic acid sequences can also be modified using many means known in the art.

The probe is an oligonucleotide capable of binding to a target nucleic acid in a base-specific manner as a nucleic acid strand partially or completely complementary to the target nucleic acid. Preferably, it is an oligonucleotide that is completely complementary to the target nucleic acid. The probe includes not only nucleic acid but also any nucleic acid derivative known in the art which is capable of complementary binding including a peptide nucleic acid. The binding of the probe to the target nucleic acid (generally, also referred to as hybridization) occurs in a sequence-dependent manner and can be performed under various conditions. Generally, the hybridization reaction occurs at a temperature about 5 ° C below the Tm for a particular sequence at a specific ionic strength and pH. The Tm means that 50% of the probe complementary to the target sequence is bound to the target sequence. An example of the hybridization reaction conditions may be a pH 7.0 to 8.3, 0.01 to 1.0 M Na + ion concentration. In addition, in order to enhance the specificity of the target nucleic acid and the probe, it is preferable to carry out the conditions for destabilizing the binding between the target nucleic acid and the probe, for example, by avoiding the presence of a high temperature, a high concentration of a destabilizer (for example, formamide) . The length of the probe is capable of specifically binding to the target nucleic acid sequence, and includes polynucleotides of any length. For example, the length of the probe may be 10 to 200 nucleotides, 10 to 150 nucleotides, 10 to 100 nucleotides, 10 to 50 nucleotides, or a single strand length of a full-length gene, but is not limited thereto. The probe may be labeled with a detectable label. The detectable label is labeled with, but not limited to, the fluorescent material FAM at the 5'-end of SYBR Green I or various probes (TaqMan Probe) and the TAMRA quencher at the 3'-end. A fluorescent label such as Cy3 or Cy5, a radioactive substance label, an enzyme for converting a substrate into a coloring material, and the like, but the present invention is not limited to these examples.

The probe or probe set may be immobilized on a microarray.

The biomarker in the sample is hybridized with the probe in the microarray, and its presence and concentration can be measured by measuring the hybridization result. During the hybridization process, the target nucleic acid may be labeled with a detectable label.

The antibody refers to a protein capable of specifically binding to an antigen epitope of an antigen, and includes a polyclonal antibody, a monoclonal antibody, and a recombinant antibody. The antibody can be easily prepared using techniques well known in the art. Specifically, the polyclonal antibody can be produced by a method well known in the art for obtaining sera containing an antibody by injecting an animal with a protein antigen encoded by the marker gene of the present invention and collecting it from an animal. Such polyclonal antibodies can be prepared from a variety of animal species hosts such as goats, rabbits, sheep, monkeys, horses, pigs, cows, dogs and the like. The antibody comprises a functional fragment of an antibody molecule as well as a complete form having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule refers to a fragment having an antigen-binding function, and includes Fab, F (ab ') 2, F (ab') 2 and Fv.

The "Yeonsan Ogye" of the present invention is a traditional breed distinguished from Japanese oysters, and is a species listed as Natural Monument No. 265 under the name of the Five Kingdoms of the Huaxia. It is mainly raised in Hwawari of Namsan-si, Nonsan-si, Chungcheongnam-do. The body is black, and the bone and eyes are all black.

The gene of the present invention can be obtained without limitation from various biological samples from which a gene can be obtained, but it can be characterized in that it is extracted from an liver sample in one embodiment of the present invention.

The present invention also provides a kit for identifying a breed of chicken comprising the composition for discriminating the breed of chicken.

The kit may further comprise one or more other components, solutions or devices suitable for analyzing the amount of gene expression or expression pattern, or the method for analyzing the amount or presence pattern of a protein in addition to nucleic acid or antibody. For example, when the kit is a kit for detecting the expression level or expression pattern of a gene, it may be a kit containing essential components necessary for performing RT-PCR. The RT-PCR kit may include a biomarker gene (DNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor (RNase inhibitor), and the like, depending on the specific embodiment, DEPC water, sterilized water, primer pairs specific for genes used as a quantitative control, and the like. On the other hand, if the kit is a kit for detecting the amount or presence pattern of a protein, the kit may be, for example, a kit containing essential components necessary for performing an ELISA, (E.g., an antibody conjugated with an antibody) and its substrate, and an antibody specific for a quantitative control protein, and the like, for example, . According to a specific embodiment, the kit may comprise a DNA microarray or a protein microarray.

The gene expression amount or expression pattern can be detected by a conventional biochemical analysis method for confirming not only the DNA microarray but also the amount or pattern of mRNA produced due to transcription of the gene. RT-PCR, competitive RT-PCR, real-time RT-PCR, and RNase protection assay were used to confirm the amount and pattern of these mRNAs. , Northern blot, and the like, and any method conventionally performed in the art may be used.

Preferably, the expression pattern of the gene biomarker in the present invention can be confirmed by a microarray method, more preferably a DNA microarray method.

The invention also relates to a nucleic acid sequence of a biomarker consisting of one or more genes as described above; And a microarray chip for discriminating the breed of chicken in which a complementary nucleic acid sequence is integrated.

The present invention also relates to a method for producing a chicken, comprising: 1) obtaining a gene sample from a chicken; 2) comparing the degree of expression of one or more genes selected from SEQ ID NO: 4 to SEQ ID NO: 8 in the sample of the first step; The present invention provides a method for distinguishing a chicken breed.

The method can classify chicken breeds with high accuracy using genetic information, classify chickens with a high degree of accuracy in distinguishing breeds of chickens that are difficult to classify by naked eyes, and breeds of chickens in hybrid chickens There are advantages to be able to.

In the present invention, when the expression of SNORA5 (SEQ ID NO: 4), CRLF2 (SEQ ID NO: 5), DETYO1 (SEQ ID NO: 6) and DETYO2 (SEQ ID NO: 7) among the genes is increased, .

When the expression of MIR1803 (SEQ ID NO: 8) in the gene is decreased, the chicken can be judged to be the fifth ovine.

The genetic markers of SEQ ID NOS: 4 to 8 mentioned above may be used singly, but they provide the ability to discriminate the breed more accurately than when used in combination of two or more kinds, and have an advantage of discriminating chickens of various kinds at one time .

It is also within the scope of the present invention that the above-described contents of the present invention are applied mutatis mutandis to each other, and those skilled in the art will make appropriate changes. Hereinafter, the present invention will be described in detail with reference to Experimental Examples and Examples, but the scope of the present invention is not limited to the following Experimental Examples and Examples.

Experimental Example  One. Traditional chicken  Obtain Derived RNA

Four broiler chickens were fed with conventional broiler chicks, Hwangbong, Hwangseong chicken, and sage broods. The broods were artificially hatched and grown according to the standard protocol of Seoul National University. Liver tissues were collected from 13 - week - old hens with active metabolism and growth, and the collected liver tissues were stored at - 80 ℃. Subsequently, transcripts (total RNA) were isolated and purified from the liver tissues collected using TRIzol reagent (Thermo Scientific) according to the protocol recommended by the manufacturer. More specifically, the following steps were carried out to separate and purify the transcript.

In step 1, tissue homogenization was performed by disrupting liver tissue contained in the TRIzol reagent with a tissue homogenizer by physical force. In step 2, the TRIzol reagent containing crushed liver tissue was treated with chloroform as a phase separation step, and the RNA layer was separated by centrifugation. In step 3, 100% isopropanol was treated with RNA precipitation and washing to extract total RNA, and impurities were removed by treatment with 70% (v / v) ethanol (EtOH).

DNase I was treated with DNase I to remove all remaining DNA, and then treated with 70% (v / v) ethanol to remove impurities. Finally, pure RNA was finally purified.

Example  One.

1.1 RNA Transpositional  Expression analysis

The purified transcripts obtained in Experimental Example 1 were measured for absorbance at a wavelength of 260 nm using a Nanodrop (Thermo Scientific) instrument and their amounts and pure water numbers were obtained. Based on this, 500 ng of formaldehyde agarose, FA) gel, and the amount and quality thereof were confirmed. Transcripts were hybridized to the Affymetrix GeneChip Chicken Gene 1.0 ST Array and their expression was measured. Affymetrix® WT PLUS Reagent Kit was used according to the method described by Affymetrix.

More specifically, 100 ng of the pure RNA-containing transcript was mixed with the Poly-A RNA Control Diluant, the Fist-Strand Buffer Mix and the First-Strand Enzyme Mix provided in the Affymetrix WT PLUS Reagent Kit for 1 hour at 42 ° C, 1-hour incubation at 4 ° C for 2 minutes to obtain first-strand cDNA. Second-strand Buffer Mix and Second-Strand Enzyme Mix are mixed and incubated at 16 ° C for 1 hour, 65 ° C for 10 minutes, and 4 ° C for 2 minutes. The IVT Buffer Mix and the IVT Enzyme Mix were mixed and incubated at 40 ° C for 16 hours to in vitro transcription reaction to amplify cRNA. The cRNA samples were then extracted by treatment with purified beads and washed with 80% (v / v) ethanol to obtain pure cRNA only. Second-cycle ss-cDNA buffer and Second-Cycle ss-cDNA enzyme were added to 15 μg of the obtained pure cRNA, and the mixture was incubated at 25 ° C. for 10 minutes, 42 ° C. for 90 minutes, and 70 ° C. for 10 minutes. Cycle Single-stranded cDNA was prepared. After that, RNase H was added and the cRNA template was removed by allowing to stand for 45 minutes at 37 ° C, 5 minutes at 95 ° C, and 2 minutes at 4 ° C. Second-Cycle ss-cDNA was purified on beads, mixed with 100% (v / v) ethanol, and washed with 80% (v / v) ethanol to obtain pure ss-cDNA. To 5.5 μg of ss-cDNA, 10 × cDNA Fragmentation Buffer, UDG, and APE1 are mixed and allowed to stand at 37 ° C. for 60 minutes, 93 ° C. for 2 minutes, and 4 ° C. for 2 minutes, and the ss-cDNA is cut into small pieces. Then, 5X Tdt buffer, Tdt enzyme, and DNA labeling reagent were mixed, followed by incubation at 37 ° C for 60 minutes, 70 ° C for 10 minutes, and 4 ° C for 2 minutes, and biotin labeling was performed at the 3'end of ss-cDNA. 3.5 μg of the target was injected into Affymetrix® GeneChip® Chicken Gene 1.0 ST Array with Control Oligo B2, 20X Hybridization Controls, 2X Hybridization Mix and DMSO and hybridized at 45 ° C for 17 hours.

The hybridized array was stained with Affymetrix® GeneChip® Fluidics 450 using Stain Cocktail 1, Stain Cocktail 2, and finally stained with Array Holding Buffer to perform array image scanning on the GeneChip® Scanner 3000. The post-hybridization procedure was performed using the Affymetrix® GeneChip® Commend Console (AGCC) Portal, which controls the Affymetrix System. The scanned array image was converted into a CEL file based on the automatically generated grid in the initial DAT file and a quantized normalized CHP file was generated using the robust multi-array (RMA) method in the Affymetrix® GeneChip® Expression Console. The result of the array was evaluated using this CHP file.

The results of the array evaluation show whether the pos vs neg auc value is 0.8 or more, the median of the RLE box plot is 0, and the pm_mean, positive control, bacterial spikes, poly A spikes, mad_residual_mean, rle_mean value, and so on.

The transcript expression database was constructed through the above hybridization process, and the constructed database contained a total of 18,308 gene expressions.

1.2 Marker  Selection of genes

The ANOVA p - value of each gene was obtained using the transcript expression database obtained in 1.1, and the FDR value of each gene was obtained and only genes with a value of <0.05 were selected. Of the selected genes, only Tukey 's post - test results showed that genes with different expression in only one line of the five lines, Hwangbong, Hoengseong chicken, and sage black were selected again. Fold change calculation was performed using the selected gene as described above, and genes with an absolute value of> 1.5 were selected again. Finally, a gene that can be used as a computational marker marker gene was derived as a native selection marker gene. The derived marker gene and the control gene, white lag horn-related gene are shown in Table 1.

[Table 1]

1.3. Selection Marker  Used Math five  Breed distinction

Whether or not the expression of each gene marker shows different expression characteristics from that of white leggong in order to determine whether the selected genetic markers can distinguish the five genes of the conventional chicken from the white regulon which can be used as a standard control group Respectively. The expression levels of the eight genes in Table 1 were compared and analyzed in a total of three individuals belonging to the white regrowth (WL) and the five genes of the five genes from the five genes of the five genes in the five genes of the five genes. The expression values shown in Table 2 are expressed as log2 scale. Table 3 shows the FC value, ANOVA p-value, FDR, Tukey post test value of 8 marker genes.

[Table 2]

[Table 3]

As shown in Tables 2 and 3, the markers in which the expression of RNA significantly increases or decreases in each of the 5 's were confirmed, and it was confirmed that the five genes could be accurately separated at the gene level. More specifically, the expression of SNORA5 (SEQ ID NO: 4), CRLF2 (SEQ ID NO: 5), DETYO1 (SEQ ID NO: 6) and DETYO2 (SEQ ID NO: 7) RNA specifically increased, and MIR1803 ) Was specifically reduced in the expression of the RNA.

When the CRLF2 gene was used as a marker, it was confirmed whether or not it was possible to specifically discriminate the five haplotypes from the white leggings, the sagittal blacks, the Hoengseong broiler chicks, and the hwangbong. The expression of the CRLF2 gene in each chicken variety was confirmed by the following Table 4 1.

 [Table 4]

As shown in Table 4 and FIG. 1, the expression of the CRLF2 gene was overexpressed specifically in the P5 genome, and thus it was confirmed that the P5 genome could be discriminated from various exotic species and native species chickens using this marker.

Since the markers whose expression is specifically changed in various mental processes have been identified as described above, it is possible to genetically distinguish the mature mushroom species by confirming the increase or decrease of the genetic markers.

<110> Korea University Research and Business Foundation <120> Composition for discrimination of Yeonsan Ogye <130> KOU1-56 <160> 8 <170> Kopatentin 2.0 <210> 1 <211> 88 <212> DNA <213> Gallus gallus <400> 1 ttgagaagta cttgtttgta ccaaaagatg atgagttgct gatgtgcttt gtcgaggtaa 60 cgctgtttcc tgggtgcagt atcgctta 88 <210> 2 <211> 872 <212> DNA <213> Gallus gallus <400> 2 tggttggaat ctttgcagga tttggtcttc tacttttggt ggcctcccct ttccttttac 60 ttgctacacc atttgttctg tgctgcaagt gcaaatgtaa taaaggagat gacgaccctc 120 tacctactta gatgactttc tcctcataca ttatgacaaa ggaccttgtc gaaataagct 180 gggacactcc agggcatctg ttatctggca cagaacacag tccatgttta aaatggaaga 240 agacgatgct ccaatcaagc gctgtccaaa gtgtaaagtt tacattgaac gagatgaggg 300 ttgtgcccag atgatgtgta agaactgcaa acatgccttt tgctggtact gtctggaatc 360 tctcgatagt gcttttggat ccctgtcgga cttggtgtcc atcctctact tgccaagcag 420 tttgccagct ccaggaatca agcccacaga accctcagct ggtccagtgc aaagcctgtg 480 acattgagtt ctgctctgtt tgcaaatcta attggcatcc aggtcaaggc tgtcaggaga 540 acatgccaat ctcttttctt ccaggagaaa caagttgagt gcatggttgc atcagaaatc 600 atgagaggt ataagaaact acagtttgaa agaggcctaa aacagtatgt ggaacttttg 660 atcaaagaag gtctagaaac ggcaatcagc tgccctgatg ctgcctgtcc aaagcgaggt 720 catttgcaag aaaatgagtg actacagcaa gatataggcc tacctgggac ttggtacttg 780 acccgttagt gtcctgcaag ctctgccttg gtgaatatcc tgtggagcag atgacaacaa 840 tagcacaatg ccaatgcatc ttctgtactc tg 872 <210> 3 <211> 916 <212> DNA <213> Gallus gallus <400> 3 tttgagctgc gaggtggagg ccaatgtgcg tacctgaatg gggacaggat cagctcagcc 60 ctatgccaca atgagaaatt ctgggtgtgc agcactgctg acagctacgt ccgctggaag 120 cagaaggcat taccagatta agcgatccca tcagcactga ggtcgttgca agtctgctgg 180 gcactgcaaa gccatcggta aatccttcca tctgcctctt tccagcactc ctgcacttgg 240 ctctgcatga aaacacacag atctgctcct cattaactct ccggcacttc atctgttcct 300 ctgtgcctta aagggagggt tctgtgagag tccttgaagg tcacagcccc acagaatgtt 360 cccacggtgc tgcttactgc agccagaccc gagggtgagg gtatttctgc tttcctttcg 420 ggctctgtgt gacatcgcgt tgccacaaat gtgtcacatg aaggtgccgt cactccaatt 480 catgctgcgg taccggggcc cagcaaactg ttggattggg ctgcacaggg cagaagggga 540 cgagcactgg acatgggccg atggcagcgc cttcaccaac tggctccagc gtccgtcgtg 600 ctcacctcgc ccacccttct cccacgtgtg ccccaacacc tcaatcggtt tccaagggaa 660 atgctattat ttctcggaca cggagagtga ttggaacagc agcagggagc actgccaccg 720 cctcggagct tccctggcta ccatagagac ggacgaggag atgctaggct ccagatgcag 780 tgagctgaga cagaacaggc gccgcgtgct ctgcgtggct ttgtgtgcag tgccgtgcat 840 ccttgtttcg gcgctggtgg ccgtgatagt ccccgccccg agaggcgacg acgcgcgaag 900 gagacaaaga acggca 916 <210> 4 <211> 137 <212> DNA <213> Gallus gallus <400> 4 gcagcccagt caaattcagt ctctgcctcc tccatggcga gcactggcca tccggctgcc 60 acagcaacat ctgtagtcca tgggccccac gtccccaccc atgggaccct ggggataaaa 120 tcgtgccaca gacactg 137 <210> 5 <211> 1049 <212> DNA <213> Gallus gallus <400> 5 aatggataga taaaactgat catgcagtgg tgcaaaccaa attagaatat gaagaacctg 60 aatgcatcat cagaggaagaa aaccagcaaa tagacaagaa aagtgacaat caggaaccta 120 gggaaaaaat cttcagcttt tcacaattaa ctggaaataa cagtcccaaa gcagctcaca 180 atgcctgcct ggtgccaaca ttcaataata tggtttcctt tgctggcctc cagaatgtag 240 tgaatgatga cacgtatgtg atgttacttc aaaaatctgt catgcctact ataccagatc 300 caaaatatgt atttactgat ctcttcaaca atcataatgg aaacttccag ttcatgtgat 360 gatgaaataa aaacccagtc aaacacagta attcttttaa gttgtttgat gacactactt 420 ctaatggtat ttatcctctt gattcttctg tgtaaatggc agagccagaa cttctaagtg 480 ttgcagagtt ggagagcaag gctttgatcc aaggaaatgc tattcttttc gggtcagact 540 gaagaggata gtaccctctt gcaacgtagt tgattacagc agtgactggg aagcagaaac 600 attttggatg aatagcacat tattagaaag cccaatcgac cagaaaatgt gaccttcttc 660 tggaaggatg acactgttac tgttacatgt aataaaccag aaaaaggtgt gaagtgcttg 720 agacttgagc ttcaatacaa aagcaaatat gacaaaggat ggcaatttga agaaggcaag 780 caaaaatctt ggaaaacatg tcccacatat ctactgacac aaggctataa tacaggatgt 840 ctttttaaga cagatggata cacccttgct ttctctatca tgaacaccaa tggaagtgaa 900 gagctttttt ctaaaagatt gaaatctgat ttctacagtc agaaagatgc catcaacacc 960 acaataatca atttcaataa tgagaaaatg cagatcacgt gggcaacaag caaactattc 1020 tctggcaaga atgtgtcatt ttattatac 1049 <210> 6 <211> 154 <212> DNA <213> Gallus gallus <400> 6 tacatccggt actcccagat ctgcgctcag gttgtcagag cggcgatgaa gcctcagtac 60 aaagctgagg cggagagggc ggcaacggcg tccgtgaaga ttgtgaagac caagaaggag 120 tagtggtggc gtactggcgg caggccggac tcag 154 <210> 7 <211> 566 <212> DNA <213> Gallus gallus <400> 7 atacatgagt atctgacaga taacctttga aatggaggca cagaagcatc ctggtctgac 60 agctgcacca gcagaccagc cacttccaac ctcttctctt cagcctgcac agggtccttc 120 agctctctgc caaagcagtg tggttcctcc acaggaacaa gcactgtctc aacctgtgta 180 tctaaaagcc ctcaccatcc ccctgtacca gcctgtccag ccaggatgct ttcagccaag 240 cagtcagcta gtggctggca ggagctgtgg aaacctggat agcagtaaca tacctctgat 300 cctgaaccca cttgttcctt ccgaaggcac agatcagccg cagtcagtgt ttcagaaaca 360 acttgggcaa actctaacat tgaacatagt gagcacttta ccagttttat catccccaaa 420 ttcttctgcc aacgcaccca ttggaagccc aggaaaatca aaaaatgctg ggaaatatat 480 ctgtaagcac tgcggtcgag actgtttgaa gccaagtgtc cttgagaaac atatccgctt 540 ttgctgaaat gtgtgcctca ataaca 566 <210> 8 <211> 88 <212> DNA <213> Gallus gallus <400> 8 gacttagggc atgatccaga gcccattgta gtcaatggga gtctttttcc attgacttca 60 gcggggtttg gatcaggcct ttacttcc 88

Claims (8)

A biomarker consisting of one or more genes selected from SEQ ID NOS: 4 to 8, a nucleic acid or oligonucleotide specific to said gene, or an antibody specific to a protein encoded by said gene. The composition for discriminating chicken varieties according to claim 1, wherein the chicken is a five-membered group. The composition for discriminating chicken breeds according to claim 1, wherein the gene is extracted from an liver sample. A kit for identifying a breed of chicken, comprising the composition of any one of claims 1 to 3. Wherein at least one gene selected from SEQ ID NOS: 4 to 8 or a nucleic acid sequence complementary to said gene is integrated. 1) obtaining a gene sample from a chicken;
2) comparing the degree of expression of one or more genes selected from SEQ ID NO: 4 to SEQ ID NO: 8 in the sample of the first step; &Lt; / RTI &gt; 7. The method according to claim 6, wherein when the expression of SNORA5 (SEQ ID NO: 4), CRLF2 (SEQ ID NO: 5), DETYO1 (SEQ ID NO: 6) and DETYO2 Wherein the chicken breed is selected from the group consisting of: 7. The method according to claim 6, wherein when the expression of MIR1803 (SEQ ID NO: 8) in the gene is decreased, the chicken is determined to be a fifth ovine.

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