KR101686436B1 - A composition for detecting intramuscular fat tissue in cow and detecting method using the same - Google Patents

Abstract

The present invention relates to a composition for detecting intramuscular fatty tissue and a method of detecting the same. More particularly, the present invention relates to a composition for detecting intramuscular fatty tissue, The expression of one or more genes selected from the group consisting of MYLPF, PGK1, ACTN3, GAPDH, CA3, PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, OGN, YWHAG, FABP4, HAUS1 and LDH A composition for detecting intramuscular adipose tissue, a kit, and a detection method. By using the present invention, it is possible to detect small intramuscular fat tissue-specific expression proteins, and thus high-quality beef with excellent marbling can be quickly and accurately selected.

Description

Technical Field [0001] The present invention relates to a composition for detecting intramuscular fat tissue and a method for detecting the same using the same.

TECHNICAL FIELD The present invention relates to a composition for detecting intramuscular fat tissue and a detection method using the composition, and more particularly, to a composition capable of rapidly and accurately selecting a high-quality animal having excellent marbling because a specific expression protein specific to intramuscular fat tissue can be detected , A kit and a detection method.

The improvement of Korean beef cattle was selected by combining the current surveillance and the later surveys. In this test, the growth ability of the individual was assessed, and the candidate sows were selected. The growth and conductance of the offspring, , And a two - way test system is being conducted to select certified sushi from candidates. However, in the present test, the trait to evaluate the hereditary ability of an individual is mainly assessed only for growth, but the meat quality data can not be measured. Therefore, it is difficult to accurately evaluate the meat quality of the subject because it is evaluated based on the breeding value of the parents. Therefore, applying genetic markers which can estimate the genetic ability of meat quality will improve the speed and accuracy of Hanwoo supplementation if selected at the time of the test. Since the recent Korea-US Free Trade Agreement (FTA) has been concluded, it is very important to increase the meat weight and quality of Korean beef meat following the opening of cheap US beef imports. Especially, to diversify the quality of imported beef and improve its competitiveness, diverse measures should be sought including the use of advanced science and technology for the production of high quality meat.

The carcass price of Hanwoo meat is evaluated by the grade of meat and meat quality. The meat quality is judged on the basis of the carcass, the thickness of the backfat, and the cross sectional area of the carcass, and meat quality is judged by the degree of muscle fat, meat color, local color, texture and maturity Livestock Classification Office, 2004). The degree of intramuscular fat is closely related to the year, juiciness and flavor, which are the criteria of sensory characteristics of beef, so it is very important to increase intramuscular fat content for high quality meat production. Genetic information on the economic performance of Hanwoo at the DNA level provides important information to early breeders as well as producers to select the best Hanwoo. Such DNA information can be used to improve the accuracy of selection by applying marker-assisted selection (MAS) method for meat and meat quality, saving the less expense and time (generation interval) .

A prior art related to the present invention is Korean Patent Laid-Open Publication No. 10-2014-0085194 (July 31, 2014).

A number of patent documents are referenced and cited throughout the present invention. The disclosures of the cited patent documents are hereby incorporated by reference into the present invention in its entirety to better illustrate the state of the art to which the present invention pertains and the content of the present invention.

Korean Patent Publication No. 10-2014-0085194 (Jul.

It is an object of the present invention to provide a composition for detecting intramuscular adipose tissue which enables practical and rapid detection of intramuscular fat tissue-specific protein.

Another object of the present invention is to provide a kit comprising the composition.

It is still another object of the present invention to provide a method for detecting intramuscular fat tissue using the composition.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to an aspect of the present invention, there is provided a method for screening for a disease caused by a mutation in a gene selected from the group consisting of small genes MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, CA3, A composition capable of measuring the expression of one or more genes selected from the group consisting of DES, TPI1, ENO3, TPM1, ATP2A1, OGN, YWHAG, FABP4, HAUS1 and LDHA is provided .

According to another aspect of the present invention, a kit for detecting small intramuscular fat tissue comprising the composition can be provided.

According to another aspect of the present invention, there is provided a method for quantifying mRNA expression, comprising: extracting mRNA of each gene from two or more bovine animals, quantifying each mRNA expression amount, And

MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, CA3, PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, And determining that the intramuscular fat is low when the mRNA expression level of at least one of OGN, YWHAG, FABP4, HAUS1 and LDHA is higher than the average mRNA expression level, is provided .

In accordance with another aspect of the present invention, there is provided a method for the treatment and / or prophylaxis of diabetes, comprising administering to a subject in need thereof an effective amount of a compound selected from the group consisting of MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, Comprising increasing or decreasing the mRNA or protein expression level of at least one gene selected from the group consisting of MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, OGN, YWHAG, FABP4, HAUS1 and LDHA A method of controlling fat may be provided.

According to one embodiment of the present invention, expression of specific intestinal tissue-specific protein can be rapidly and accurately detected.

According to one embodiment of the present invention, a high-quality beef having excellent meat quality and marbling can be quickly and accurately selected.

In addition, according to one embodiment of the present invention, it is possible to develop a technique for controlling body composition including high fat production through intramuscular fat-specific fat content enhancement.

FIG. 1 shows electrophoresis results showing intramuscular fat tissue-specific protein expression according to intramuscular fat diagram according to an embodiment of the present invention.
Figure 2 shows immunoblot expressing specific expression of TPM2 protein according to an embodiment of the present invention and a bar graph thereof.

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

According to an aspect of the present invention, there is provided a method for screening for a compound of the present invention, which comprises the steps of: A composition capable of measuring the expression of one or more genes selected from the group consisting of DES, TPI1, ENO3, TPM1, ATP2A1, OGN, YWHAG, FABP4, HAUS1 and LDHA is provided .

In the present invention, the term " intramuscular fat "is edible fat. The higher the content of the intramuscular fat, the better the meat quality and marbling of the beef.

The gene used in the present invention can be obtained from a known database such as GeneBank of the National Institutes of Health.

In the present invention, "measuring the expression level of a gene" may be a measure of the level of mRNA or protein.

Any of known gene measurement or detection methods known in the art can be used to measure the expression level of a gene. The above "measurement of the expression level of the gene" can be analyzed by any conventionally known method including RT-PCR, competitive RT-PCR, real-time RT-PCR, RNase protection assay, Northern blotting, DNA microarray and the like . Preferably, mRNA isolated from the biological sample or cDNA derived therefrom is hybridized on a microarray in which a probe specific to one or more marker genes selected from the group consisting of the genes is immobilized, and the resulting degree of hybridization is measured . The hybridization degree can be measured by any measurement method known in the art such as fluorescence measurement and electrical measurement. In this case, the probe or the target nucleic acid may be labeled with a detectable appropriate label. Here, the cDNA may be directly amplified by RT-PCR using a pair of sense and antisense primers targeting at least one marker gene selected from the group consisting of the genes as primers.

In the present invention, the above-mentioned "measurement of the level of the protein" may be performed using any of known protein measurement or detection methods. For example, an assay method using an antibody that specifically binds to a protein expressed from one or more marker genes selected from the group consisting of the genes may be used. Methods for analyzing proteins using antibodies include Western blotting, ELISA, radioimmunoassay, radial immunodiffusion, Oucheronin immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assays, complement fixation assays, FACS But are not limited to these examples. Such ELISAs include direct ELISA, indirect ELISA, direct sandwich ELISA, indirect sandwich ELISA, and the like. Western blotting refers to separation of whole proteins, electrophoresis, separation of proteins according to their size, transfer to a nitrocellulose membrane, reaction with the antibody, and quantification of the amount of the produced antigen-antibody complex by using labeled antibodies It is a way to confirm. In addition, methods for measuring protein levels include methods using enzymes, substrates, coenzymes, and ligands that specifically bind to target proteins.

In the present invention, the "agent" may further include a primer set capable of amplifying the gene, a reagent necessary for hybridization with the gene or the nucleic acid expression product expressed therefrom. In addition, the preparation may further comprise a buffer, a solvent, or the like serving as a medium for the reaction.

In one embodiment, the agent for measuring the expression level of the gene is provided with a composition for detecting intramuscular adipose tissue comprising at least one of primers specifically binding to the gene represented by SEQ ID NOS: 1 to 60 (See Table 4).

As used herein, the term "primer " refers to a single-stranded oligonucleotide sequence complementary to a nucleic acid strand to be copied, and may serve as a starting point for the synthesis of a primer extension product. The length and sequence of the primer should allow the synthesis of the extension product to begin. The primer preferably has a length of 15 to 30 base pairs.

The oligonucleotide used as the primer may also comprise a nucleotide analogue such as phosphorothioate, alkylphosphorothioate or peptide nucleic acid or an intercalating agent.

According to another aspect of the present invention, there is provided a kit for detecting intramuscular fat tissue comprising the composition.

In the present invention, when applied to a PCR amplification process, a "kit" may optionally contain reagents necessary for PCR amplification, such as buffers, DNA polymerases (e.g., Thermus aquaticus (Taq), Thermus thermophilus , Thermisflavus, Thermococcus literalis or Pyrococcus furiosus (Pfu)), DNA polymerase joins and dNTPs. The kit may be made from a number of separate packaging or compartments containing the reagent components described above.

In one embodiment, the kit may be an RT-PCR kit, a microarray chip kit, or a protein chip kit.

The RT-PCR kit may comprise a respective pair of primers specific for the marker gene, and may include other test tubes or other appropriate containers, reaction buffers (varying in pH and magnesium concentration), deoxynucleotides (dNTPs), Taq Enzymes such as polymerase and reverse transcriptase, DNAse, RNAse inhibitor DEPC-water, sterile water, and the like.

The protein chip kit may be a kit in which one or more antibodies against a marker are arranged at predetermined positions on a substrate and fixed at high density. In the method using a protein chip, a protein is separated from a sample, and a separated protein is hybridized with a protein chip to form an antigen-antibody complex, and the presence or the expression level of the protein can be confirmed by reading it.

According to another aspect of the present invention, there is provided a method for quantifying mRNA expression, comprising the steps of: extracting mRNA of each gene from two or more bovines, quantifying each mRNA expression amount, And

MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, CA3, PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, And determining that the intramuscular fat is low when the mRNA expression level of at least one of OGN, YWHAG, FABP4, HAUS1 and LDHA is higher than the average mRNA expression level, is provided .

In one embodiment of the present invention, the step of extracting proteins from two or more bovines, quantifying each protein expression amount, and obtaining an average protein expression amount; And

MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, CA3, PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, And determining that the intramuscular fat is low when the amount of the protein expressed from at least one gene of OGN, YWHAG, FABP4, HAUS1 and LDHA is higher than the average protein expression amount, Can be provided.

In another aspect of the present invention there is provided a method for the treatment and / or prophylaxis of endometrial hyperplasia comprising administering to a subject in need thereof a therapeutically effective amount of at least one compound selected from the group consisting of MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, Comprising increasing or decreasing the expression level of mRNA or protein of at least one gene selected from the group consisting of PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, OGN, YWHAG, FABP4, HAUS1 and LDHA A method of controlling intramuscular fat may be provided.

Example  1: Animals and Samples

Abdominal, subcutaneous and intramuscular fat of Hanwoo steer, hydrogen and cow were collected and stored in liquid nitrogen and stored in freezer at -80 ℃. Site-specific proteins were selected based on the results of 1D gel and LC-MS / MS identification of each sample. Table 1 below is the disclosure material used for the analysis. (7.00 ± 0.94), intermediate group (4.00 ± 0.58) and lower group (1.00 ± 0.00) in 21 Hanwoo (cow, hydrogen and steer) groups. The individual number, slaughter age and trait characteristics of the samples used in the analysis can be found in Table 1. Generally, the higher the intramuscular fat level, the better the meat quality and marbling.

Example  2: Protein Isolation

Prepare lysis buffer to prepare the lysis buffer [8M Urea 4.8g + 2M Thio-urea 1.52g + 40mM Tris 0.048g + 100mM DTT 0.154g + 2% Chaps 0.2g -> Water (ultra pure grade) mass up 10mL] And 2 ml of lysis buffer was added per mg of adipose tissue. Homogenized using a sonicator and centrifuged at 5,000 rpm, 4 ° C for 30 minutes to separate the layers. After removing the layer, the upper layer of fat layer was removed and the lower layer was transferred to a new 1.5 mL tube. And centrifuged again at 12,000 rpm, 4 캜 for 30 minutes. The layered product was transferred to a new 1.5 mL tube. The concentration was determined by measuring the absorbance at 595 nm using the Bradford method.

Example  3: Primary electrophoresis

The prepared protein samples were electrophoresed with 15% polyacrylamide gel to confirm the separation pattern of protein bands.

Example  4: LC - MS - MS Protein identification through

Each protein DB was identified and the spectral count was quantified using label-free. Relative quantification using spectral count was indirectly measured by using the spectral count (number of identified peptides) from protein identification.

Example  5: Identification of proteins

The cold Pro-PREP solution was added to the adipose tissue, homogenized using a homogenizer, the proteins were separated and quantitated by BCA assay. The homogenized protein (50 ug) was separated by SDS-PAGE, transferred to PVDF membrane, and blocked with 1xPBS / 0.1% Tween 20 and 5% nonfat dry milk powder. Immunoblotting is performed using a primary antibody (diluted factor = 1: 100 ~ 1: 1000) for the candidate protein. The secondary antibody of anti-rabbit or anti-mouse antibody reacting with Blot was bound to horseradish peroxidase (Abcam) and activated by chemiluminescence reagent (Millipore, Billerica, MA). Blots were measured using an EZ-capture II chemiluminescence imaging system and images were quantified using capture system analyzer version 2.00 software. Band thicknesses were normalized using α-tubulin.

Experiment result

1) Primary electrophoresis

Electrophoresis (1D) was performed according to the intrauterine fat level (low fat vs. high fat), one of the economic traits, using Hanwoo intramuscular fat (Fig. 1). For the first electrophoresis, protein samples separated and purified were electrophoresed with 15% polyacrylamide gel and their protein bands were identified by silver staining.

2) LS - Mass - Mass Of the expressed protein Profiling

Table 2 shows the proteins specifically expressed by marbling in the adipose tissue of Hanwoo. Overall, about 40 proteins were observed.

3) LC - Mass - Mass Hanwoo through Intramuscular fat  Identification of specific expression proteins

Table 3 shows that there is a strong negative correlation with the degree of marbling, except for some genes among 40 proteins specifically expressed in adipose tissue. GLM method was used statistically using SAS program. Correlation between protein expression and backfat thickness was also used. Table 4 shows primer sets (SEQ ID NOS: 1 to 60) used for amplifying the genes in Table 3 above.

4) Western blot  Hanwoo through analysis Intramuscular fat  Identification of specific expression proteins

In this study, it is suggested that nutrient metabolism and regulation and signal transduction of metabolites are influential to produce high quality Korean beef meat. Based on this concept, it is possible to obtain the source material for controlling the accumulation of fat in the fat tissue region in the Korean beef cattle through the identification of protein specifically expressed in the muscle fat tissue in relation to the intramuscular fat level of the conductor, Respectively. Tropomyosin alpha-2 chain (TPM2) protein showed a strong negative correlation with intramuscular fat tissue, but showed a difference in sex-specific expression (Fig. 2).

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.

<110> RURAL DEVELOPMENT ADMINISTRATION <120> A composition for detecting intramuscular fat tissue in cow and          detecting method using the same <130> NPF-27065 <160> 60 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 1 actgggacga catggagaag 20 <210> 2 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 2 gtcatcttct cgcggttagc 20 <210> 3 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 3 ttcaatgtcc cagccatgta 20 <210> 4 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 4 catctccaga gtccagcaca 20 <210> 5 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 5 gctatgagga ttggctgctc 20 <210> 6 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 6 tagtccctct ggctcagcat 20 <210> 7 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 7 gcatcccttc gtgaaaccta 20 <210> 8 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 8 agggaggtct gggctatcat 20 <210> 9 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 9 gcccaggaag aatacgtcaa 20 <210> 10 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 10 gccttaccac ttggggtgta 20 <210> 11 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 11 aatgggtatc gctggaactg 20 <210> 12 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 12 cgtccccaca taactgcttt 20 <210> 13 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 13 gacgtgagaa aggcgagttc 20 <210> 14 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 14 aggacgggtt gaagttgttg 20 <210> 15 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 15 gagatccgac acctcaagga 20 <210> 16 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 16 gtccaaggcc atcttgacat 20 <210> 17 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 17 acagtcacca accccaaaag 20 <210> 18 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 18 attcagtcac cgagccaatc 20 <210> 19 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 19 ggatggaaaa tcaaccacca 20 <210> 20 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 20 gtggcagtga caccattcat 20 <210> 21 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 21 acccagaaga ctgtggatgg 20 <210> 22 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 22 ttgagctcag ggatgacctt 20 <210> 23 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 23 ccagcactgg ttccaggtat 20 <210> 24 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 24 tggtgcgaaa cagatcagag 20 <210> 25 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 25 tggcagcctt ttccttagaa 20 <210> 26 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 26 acgctggacc aaattcagac 20 <210> 27 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 27 gacaactcct ctcgctttgg 20 <210> 28 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 28 gccttcagct ggaaagtgac 20 <210> 29 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 29 tggtgactca gaggcaagtg 20 <210> 30 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 30 caccgtctgg aaagaagagc 20 <210> 31 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 31 ccaagttccg aaaagctcag 20 <210> 32 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 32 tgaagtctcg ggtctttgct 20 <210> 33 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 33 ttcctctcgc tttgtgaggt 20 <210> 34 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 34 gccttcagct ggaaagtgac 20 <210> 35 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 35 ccacccaagt tcgacaagat 20 <210> 36 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 36 ttgatggtga cgcagaagag 20 <210> 37 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 37 ggaaagatcc tgcgtatcca 20 <210> 38 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 38 ggttcctctt cagctgttcg 20 <210> 39 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 39 accccagcaa tgaagagatg 20 <210> 40 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 40 gccttccttg tcgaagacac 20 <210> 41 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 41 ccatgatgaa ggaagccagt 20 <210> 42 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 42 aggctccagt gatcacatcc 20 <210> 43 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 43 ctgcaaggct aatgacacca 20 <210> 44 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 44 gatgttttcc caggatgacg 20 <210> 45 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 45 ccagaagtgg agaaggcttg 20 <210> 46 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 46 gcatcttttc ccttcccttc 20 <210> 47 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 47 gcatcaatat ggcacacctg 20 <210> 48 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 48 atgaggctcc agctcgtaga 20 <210> 49 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 49 agaaggtcgt tttcgagcaa 20 <210> 50 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 50 aggctcatag gccaagacaa 20 <210> 51 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 51 cccgtaagct ggtcatcatt 20 <210> 52 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 52 cctgagcctc cagtgatttc 20 <210> 53 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 53 aaagatggag ctgcaggaga 20 <210> 54 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 54 ctcatatttg cggtccgaat 20 <210> 55 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 55 cgctcaaagg gactaacgag 20 <210> 56 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 56 acgagccatc tcttccttca 20 <210> 57 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 57 agacatctgc cgacggtaac 20 <210> 58 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 58 ctgacatacg gcctccaact 20 <210> 59 <211> 22 <212> DNA <213> Bos taurus coreanae <400> 59 cgcatctgta ccccattcta tc 22 <210> 60 <211> 20 <212> DNA <213> Bos taurus coreanae <400> 60 catctccaga gtccagcaca 20

Claims (7)

(NM_001034502), Fructose-bisphosphate aldolase (NM_001101915), ACTN3 (Alpha-actinin-3; NM_001076157), OGN (Mimecan NM_173946), YWHAG (14-3-3 protein gamma; AF043737 ), TPM1 (Tropomyosin alpha-1 chain NM_001013590), FABP4 (Fatty acid-binding protein, adipocyte NM_174314), ACTA1 (Actin, alpha skeletal muscle NM_174225), MYH2 (Myosin- 2, NM_001166227), MYH1 1, NM_174117), MYH4 (Uncharacterized protein (Fragment); XM_615306), VIM (Vimentin; NM_173969), TPM2 (Tropomyosin beta chain; NM_001010995) and DES (Desmin; NM_001081575) Wherein the composition is used for detecting intramuscular fatness.
The method according to claim 1,
L-lactate dehydrogenase A chain (NM_174099), MYH8 (Uncharacterized protein NM_001206174), TPI1 (Triosephosphate isomerase NM_001013589), MYH7 (NM_001013589), Glyceraldehyde-3-phosphate dehydrogenase (Myosin-7; NM_174727), MYH3 (Uncharacterized protein; NM_001101835), Myosin light chain 1/3, Skeletal muscle isoform NM_001079578, Actin, cytoplasmic 1, PYGM, Phosphorylase NM_175786, ATP2A1 (Transmission ATPase; AF332982), Carbonic anhydrase 3 (NM_001034437), ALB (Serum albumin; AF542068), ENO3 (Beta-enolase NM_001034702), HAUS1 (HAUS augmin-like complex submit 1, NM_001038557), TPM1 -1 chain NM_001013590), and MYLPF (Myosin regulatory light chain 2; NM_001075647) gene.
3. The method of claim 2,
Wherein the agent for measuring the expression level of the gene comprises primers specifically binding to the gene represented by SEQ ID NOS: 1 to 60.
A kit for detecting intra-muscular fatness comprising the composition of any one of claims 1 to 3.
5. The method of claim 4,
Wherein the kit is an RT-PCR kit, a microarray chip kit, or a protein chip kit.
Extracting mRNA of each gene from two or more bovines, quantifying each mRNA expression amount, and obtaining an average mRNA expression amount; And
MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, CA3, PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, And determining that the intramuscular fat is low when the mRNA expression level of the OGN, YWHAG, FABP4, HAUS1, and LDHA genes is higher than the average mRNA expression level.
Extracting proteins from two or more bovines, quantifying the amount of each protein expressed and obtaining an average protein expression level; And
MYH2, MYH1, MYH8, MYH4, MYH7, MYH3, ACTA1, ACTA2, ALB, ALDOA, TPM2, ACTB, VIM, MYLPF, PGK1, ACTN3, GAPDH, CA3, PYGM, MYL1, DES, TPI1, ENO3, TPM1, ATP2A1, Determining that the intracavitary fat is low when the amount of protein expressed from the gene of OGN, YWHAG, FABP4, HAUS1 and LDHA is higher than the average protein expression amount.

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