KR20080006719A - Dna marker of adipocyte-fatty acid binding protein gene related the intramuscular fat content in beef cattle - Google Patents

Dna marker of adipocyte-fatty acid binding protein gene related the intramuscular fat content in beef cattle Download PDF

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KR20080006719A
KR20080006719A KR1020060065802A KR20060065802A KR20080006719A KR 20080006719 A KR20080006719 A KR 20080006719A KR 1020060065802 A KR1020060065802 A KR 1020060065802A KR 20060065802 A KR20060065802 A KR 20060065802A KR 20080006719 A KR20080006719 A KR 20080006719A
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박응우
윤두학
이승환
조용민
오성종
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Abstract

A DNA marker is provided to determine an individual genetic trait of Korean native cattle by using genotypes of three independent loci of a FABP4(adipocyte-fatty acid binding protein) gene which has the significantly effect for carcass weight and marbling of the Korean native cattle, thereby being utilized for an early selection of a fatting beef and an accurate estimate of breeding values. A method for selecting a DNA marker of determining genetic traits comprises the steps of: (a) identifying a genotype of a linkage group detected from a Korean native cattle a fatty acid binding protein(FABP4) gene of SEQ ID : NO. 7 using the decoding of 17 SNP loci depicted at a Fig. 1; (b) after pre-denaturing the Korean native cattle FABP4 gene of SEQ ID : NO. 7 as a template using a PCR primer of SEQ ID : NOs. 1 to 6 at a temperature of 94 deg.C for 5 minutes, performing 35 times of a cycle where one minute annealing at a temperature of 58 deg.C and two-minute expanding at a temperature of 72 deg.C are done and then finally reacting it for 10 minutes at a temperature of 72 deg.C for PCR amplification; (c) after cleaving an amplification product obtained from the step(b) with an restriction enzyme such as Hpy188I, RsaI and NlaIII, subjecting it to an electrophoresis; and (d) obtaining a picture and an image of the electrophoresis product to determine an individual genotype through a restriction fragment length polymorphism analysis, a heteroduplex analysis, a single strand conformational polymorphism(SSCP) and a Taqman probe method. A DNA marker is selected by the method and constructed to recognize three loci of Hpy188I(C->G), RsaI(A->T) and NlaIII(A->G) described in the Fig.1 among variable regions of a FAB4 gene expressed in adipocyte.

Description

소 근내지방도 관련 ＦＡＢＰ4 유전자를 이용한 ＤＮＡ 마커{DNA marker of adipocyte-fatty acid binding protein gene related the intramuscular fat content in beef cattle}DNA marker of adipocyte-fatty acid binding protein gene related the intramuscular fat content in beef cattle}

도 1은 한우 FABP4(Adipocyte-Fatty Acid Binding Protein) 유전자 구조변이 부위를 보여주는 모식도이다.1 is a schematic diagram showing a region of the structural modification of Hanwoo FABP4 (Adipocyte-Fatty Acid Binding Protein).

도 2는 한우 FABP4 유전자의 염기서열을 보여주는 모식도이다.Figure 2 is a schematic diagram showing the nucleotide sequence of the Hanwoo FABP4 gene.

본 발명은 한우에서의 개체별 유전적 차이를 확인하기 위한 것으로 근내지방도, 등지방두께 및 등심지방함량 등의 육질형질에 영향을 미치는 요인에 관여하는 유전적 표지인자(genetic marker)의 개발과 그 사용 방법에 관한 것이다.The present invention is to identify genetic differences between individuals in Korean cattle, and to develop genetic markers related to factors affecting meat quality such as muscle fat, back fat thickness and sirloin content and their development. It is about how to use.

지방세포(adipocyte)에서 발현되는 지방산 결합 단백질(fatty acid binding protein, FABP4)은 동물의 지방세포에서 특이적으로 발현하는 단백질로서 유리지방산과 결합하여 지방산을 중성지방의 형태로 축적시키는 지방세포로 전달하는 역할 을 한다. 가축이 사료를 섭취한 후 사료에 존재하는 지방은 소화·분해·흡수되는 과정에서 지방산으로 전이가 일어나고, 이들 유리 지방산들이 근육세포 및 지방세포 등에 유입되어 에너지원 또는 저장형태로 축적하게 된다. 이때 유리지방산형태로 존재하는 긴사슬 지방산은 지방산 결합 단백질과 결합하여 세포내 지방산화의 주요 표적 기관인 미토콘드리아로 유입되고, 이곳에서 지방산합성체계의 기본물질인 동시에, 다른 장쇄지방산합성의 전구물질로 사용되게 된다. Fatty acid binding protein (FABP4), which is expressed in adipocytes, is a protein specifically expressed in animal fat cells that binds to free fatty acids and delivers them to fat cells that accumulate fatty acids in the form of triglycerides. To play a role. After ingestion of livestock feed, fats present in the feed are converted into fatty acids in the process of digestion, decomposition and absorption, and these free fatty acids flow into muscle cells and fat cells and accumulate as energy sources or storage forms. In this case, long-chain fatty acids in the form of free fatty acids are introduced into the mitochondria, which are the main target organs of intracellular fatty acid by binding to fatty acid binding proteins, and used here as a basic material of the fatty acid synthesis system and as a precursor of other long-chain fatty acid synthesis. Will be.

근내지방도는 쇠고기 산업에 있어서 가장 중요한 경제형질 중 하나로서, 쇠고기의 풍미, 다즙성, 연도 등을 결정한다. 따라서, 근내지방 연관 유전적 표지인자 탐색에 대한 연구가 국제적으로 활발하게 이루어지고 있다. 최근, 돼지에 있어서 심장(Heart)-지방산 결합 단백질(H-FABP)이 돼지의 근내지방도와 밀접하게 연관이 있다는 다수의 연구결과가 보고되어 있다. 특히, 소에 있어서 FABP4는 염색체 14번에 IL7 마커와 17 cM 떨어진 위치에 존재하고 있으며, 이 좌위는 근내지방도와 연관된 QTL 영역인 것으로 보고되었다.Intramuscular fat is one of the most important economic traits in the beef industry and determines the flavor, succulentity, and year of the beef. Therefore, researches on genetic markers associated with intramuscular fat have been actively conducted internationally. Recently, a number of studies have reported that heart-fatty acid binding protein (H-FABP) in pigs is closely associated with intramuscular fat in pigs. Specifically, in cattle, FABP4 is located 17 cM away from the IL7 marker on chromosome 14, and this locus is reported to be a QTL region associated with intramuscular fat.

이에 본 발명자들은 근육내 지방세포 분화시에 활발하게 활성화되는 FABP4가 유리지방산을 세포내로 전달하여 근육내 지방을 축적하는데 비교적 크게 작용하여 근내지방도를 조절하고 고기의 육질을 조절하는 것으로 판단하고 FABP4를 암호화하는 유전자의 구조변이를 찾아내어, 이들 변이가 한우의 육질 등 경제적 주요 형질에 미치는 효과를 분자유전학적 분석 및 양적유전학적 통계분석에 의하여 구명하여 DNA 표지인자를 개발하여 본 발명을 완성하였다.Accordingly, the present inventors determined that FABP4, which is actively activated during differentiation of intramuscular adipocytes, acts relatively largely to accumulate intramuscular fat by delivering free fatty acids intracellularly, thereby controlling intramuscular fat and controlling meat quality. The structural variants of the genes encoding were identified, and the effects of these variants on major economic traits such as meat quality of Korean cattle were examined by molecular genetic analysis and quantitative genetic analysis to develop DNA markers to complete the present invention.

따라서, 본 발명의 목적은 한우의 성장률, 육량 및 육질 등에 관련된 DNA 표지인자를 발굴하고 그 표지 인자를 이용하여 한우 개체들의 정확한 유전적 자질을 판단하여 선발하는 방법 및 한우의 개량에 활용하는데 사용하는 방법을 제공하는 것이다.Accordingly, an object of the present invention is to find a DNA markers related to the growth rate, meat and meat quality of Hanwoo, and to use the markers to determine and select accurate genetic qualities of Hanwoo individuals and to use them in improving Korean beef. To provide a way.

상기 목적을 달성하기 위하여, 본 발명에서는 근내지방도에 유의적인 FABP4 유전자의 17개의 변이 부위들을 검출하기 위한 DNA 표지인자를 제공하되, 이들 변이들이 3개의 연관집단으로 구성됨에 따라 3개의 연관집단을 대표할 수 있는 좌위로 엑손 첫 번째 염기를 1번으로 하여 2775번째 Hpy188I (C⇒G), 3474번째 RsaI (A⇒T) 및 3632번째 NlaIII (A⇒G) 3좌위 변이부위 및 이 좌위를 검출하기 위한 DNA 표지인자를 제공한다. In order to achieve the above object, the present invention provides a DNA marker for detecting 17 mutation sites of the FABP4 gene, which is significant for intramuscular fat, but represents three association groups as these mutations are composed of three association groups. Detecting the 2775th Hpy188I (C⇒G), the 3474th RsaI (A⇒T) and the 3632th NlaIII (A⇒G) 3rd locus with this exon 1 base as the possible locus DNA markers are provided for.

이하 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail.

먼저, 본 발명에서는 한우 FABP4 유전자에서 검출된 17개의 유전자 변이 부위를 선발한 다음 이들의 연관군을 확인하였고, 이들 중 대표적인 변이를 활용 근내지방도에 유의적인 영향을 미치는가를 찾고자 하였다.First, in the present invention, 17 gene mutation sites detected in the Hanwoo FABP4 gene were selected, and then their associated groups were identified, and the representative mutations among them were used to find out whether they significantly influence intramuscular fat.

도 1은 서열번호 7의 한우 FABP4 유전자에서 변이된 17개 부위를 각각 보여주고 있다. 이를 보다 상세히 살펴보면, 유전자의 1번 엑손 첫번째 염기를 1번으로 하였을 때, 611번째 염기인 A가 G로 치환되고, 633번째 염기인 A가 G로 치환되며, 1003번째 염기인 C가 T로 치환되고, 1055번째 염기인 T가 C로 치환되며, 1151번째 염기인 A가 G로 치환되고, 1610번째 염기가 결실 또는 삽입되며, 2207번째 염기인 G가 T로 치환되고, 2775번째 염기인 C가 G로 치환되며 , 2930번째 염기인 A가 G로 치환되고, (3191)번째 염기가 TA 반복서열 부위를 가지고 있는 Microsatellite(TA) 좌위, 3474번째 염기인 A가 T로 치환되고 , 3632번째 염기인 A가 G로 치환되며 , 3652번째 염기인 G가 C로 치환되고, 3686번째 염기인 T가 C로 치환되며, 3707번째 염기인 T가 C로 치환되고, 3955번째 염기인 T가 C로 치환되며, 4163번째 염기인 A가 T로 치환된다. Figure 1 shows each of the 17 sites mutated in the Hanwoo FABP4 gene of SEQ ID NO. In detail, when the first base of the exon 1 of the gene is 1, the 611th base A is substituted with G, the 633th base A is substituted with G, and the 1003 base C is substituted with T. 1055 base T is replaced with C, 1151 base A is substituted with G, 1610 base is deleted or inserted, 2207 base G is substituted with T, 2775 base C is substituted Substituted with G , 2930th base A is substituted with G, (3191) base is Microsatellite (TA) locus having a TA repeat region, 3474th base A is substituted with T , 3632 base A is substituted with G , 3652rd base G is replaced with C, 3686th base T is substituted with C, 3707th base T is substituted with C, and 3955th base T is substituted with C. , A, which is the 4163th base, is substituted with T.

본 발명에서는 상기 FABP4 유전자의 17개 유전자 변이 부위의 연관군들 중 각 연관을 대표하는 2775번째 Hpy188I (C⇒G), 3474번째 RsaI (A⇒T) 및 3652번째 NlaIII (A⇒G) 3좌위 변이부위가 근내지방도에 유의적임을 확인하였다. In the present invention, the 2775th Hpy188I (C⇒G), the 3474th RsaI (A⇒T) and the 3652th NlaIII (A⇒G) 3 locus representing each association among the 17 gene mutation sites of the FABP4 gene. The mutant site was found to be significant for intramuscular fat.

따라서, 본 발명에서 한우의 성장률, 육량 및 육질 등에 관련된 유전적 표지인자를 발굴하고 그 표지 인자를 이용하여 한우 개체들의 정확한 유전적 자질을 판단하여 선발하는 방법은 하기 단계들을 포함한다:Therefore, in the present invention, a method for discovering genetic markers related to growth rate, meat and meat quality of Hanwoo, and determining and selecting accurate genetic qualities of Hanwoo individuals using the marker factors includes the following steps:

1) 서열번호 7의 한우 FABP4 유전자로부터 발견되고 도 1에 도시된 17개의 SNP 좌위 판독을 이용하여 연관군의 유전자형을 판별하는 단계;1) determining the genotype of the associated group using the 17 SNP locus reads found from the Hanwoo FABP4 gene of SEQ ID NO: 7 and shown in FIG. 1;

2) 서열번호 1 내지 서열번호 6의 PCR 프라이머를 이용하여 서열번호 7의 한우 FABP4 유전자를 주형으로 94℃에서 5분간 예비변성 후, 94℃에서 1분간 변성, 58℃에서 1분간 어닐링 및 72℃에서 2분간 확장하는 사이클을 35회 실시하고, 마지 막으로 72℃에서 10분간 더 반응시켜 PCR 증폭하는 단계;2) After preliminary modification of the Hanwoo FABP4 gene of SEQ ID NO: 7 as a template for 5 minutes at 94 ° C using PCR primers of SEQ ID NO: 1 to SEQ ID NO: 6, denaturation at 94 ° C for 1 minute, annealing at 58 ° C for 1 minute, and 72 ° C. 35 cycles of expanding for 2 minutes at and finally amplifying by further reacting at 72 ° C. for 10 minutes;

상기에서 PCR 반응 과정을 살펴보면, 94℃에서 5분간 예비변성 후, 94℃에서 1분간 변성, 58℃에서 1분간 어닐링 및 72℃에서 2분간 확장하는 사이클을 35회 실시하고, 마지막으로 72℃에서 10분간 더 반응시킨다.Looking at the PCR reaction process, after 5 minutes of preliminary modification at 94 ℃, 1 cycle at 94 ℃, 1 cycle at 58 ℃ annealing and extending for 2 minutes at 72 ℃ 35 times, and finally at 72 ℃ Let it react for 10 more minutes.

3) 상기 증폭산물을 제한효소 Hpy188I, RsaI 및 NlaIII로 절단하여 전기영동을 실시하는 단계; 및3) electrophoresis by cleaving the amplified product with restriction enzymes Hpy188I, RsaI and NlaIII; And

4) 상기 전기영동 결과물에 대한 사진 및 이미지를 확보하여 개체별 유전자형을 결정하는 단계.4) determining the genotype of each individual by obtaining pictures and images of the electrophoretic results.

본 발명에 의한 상기 소 FABP4 유전자의 17개 변이들을 이용하여 연관군의 유전자형을 판별하는 것으로 대표적인 Hpy188I (C⇒G), RsaI (A⇒T) 및 NlaIII (A⇒G) 3좌위에 대한 DNA 표지인자 판별 방법으로는 제한효소 절편길이 다양성(Restriction fragment length polymorphism, RFLP) 분석, 이형접합자 분석(heteroduplex analysis), 단일 나선의 구조적 다형성(single strand conformational polymorphism, SSCP) 및 테크맨 프로브법(Taqman probe method)을 들 수 있다.DNA markers for Hpy188I (C⇒G), RsaI (A⇒T) and NlaIII (A⇒G) tridents, which are representative of genotypes of related groups using 17 variants of the bovine FABP4 gene according to the present invention Factor determination methods include restriction fragment length polymorphism (RFLP) analysis, heteroduplex analysis, single strand conformational polymorphism (SSCP), and Techman probe method (Taqman probe method). ).

이하, 실시예 및 시험예를 들어 본 발명을 보다 자세하게 설명한다. 그러한 실시예 또는 시험예들은 본 발명을 구체적으로 설명하려는 것이지, 이러한 실시예 또는 시험예에 의하여 본 발명의 권리범위가 제한되는 것은 아니다.Hereinafter, an Example and a test example are given and this invention is demonstrated in detail. Such examples or test examples are intended to illustrate the present invention in detail, but the scope of the present invention is not limited to these examples or test examples.

[실시예 1] Example 1

PCR 증폭에 사용한 FABP4 유전자 분석용 프라이머는 한우 후대검정우 24두로부터 얻은 FABP4 유전자의 전장염기서열 분석 결과를 근거로 제작하여 사용하였다. The primer for FABP4 gene analysis used for PCR amplification was prepared and used based on the result of full length nucleotide sequence analysis of FABP4 gene obtained from 24 Korean cows.

한우 FABP4 유전자 변이좌위의 연관군, 치환염기, 주변염기서열 및 확인 가능 제한효소 Association Group, Substitution Base, Peripheral Base Sequence, and Recognized Restriction Enzyme in Hanwoo FABP4 Gene 연관군Associative group 좌위Seat 치환염기Substituted base 주변염기서열Surrounding base sequence 확인가능 제한효소Identifiable restriction enzyme 1One 611611 A ⇒ GA ⇒ G ttccttRaaagtcttccttRaaagtc MseI Mse I 1One 633633 A ⇒ GA ⇒ G tttgRcttaatttgRcttaa -- 1One 10031003 C ⇒ TC ⇒ T agacttYtgactaagacttYtgacta Hpy188I Hpy 188I 1One 10551055 T ⇒ CT ⇒ C atattaYaggactatattaYaggact -- 1One 11511151 A ⇒ GA ⇒ G tcctatRtggtgctcctatRtggtgc -- 1One 16101610 IndelIndel indel Tindel T -- 1One 22072207 G ⇒ TG ⇒ T atgtggKttttgtatgtggKttttgt -- 1One 27752775 C ⇒ GC ⇒ G ttttccSaactatttttccSaactat HpyHpy 188I188I 22 29302930 A ⇒ GA ⇒ G gatgaaRtcactccgatgaaRtcactcc Tsp45I Tsp 45I MSMS 31913191 (TA)n(TA) n (TA)n(TA) n -- 22 34743474 A ⇒ TA ⇒ T gctaagWacctcagctaagWacctca RsaRsa II 22 36323632 A ⇒ GA ⇒ G aaactcRtggatgaaactcRtggatg FatI, Nla III, BssSI, CviAII Fat I, Nla III , Bss SI, Cvi AII 33 36523652 G ⇒ CG ⇒ C ggtgctSgtgagtggtgctSgtgagt BssSI, Hpy188III, BsiHKAI Bss SI, Hpy 188III, Bsi HKAI 22 36863686 T ⇒ CT ⇒ C agatttYagtgctagatttYagtgct -- 22 37073707 T ⇒ CT ⇒ C cataatYgttatccataatYgttatc Tsp509I Tsp 509I 22 39553955 T ⇒ CT ⇒ C tttgtgYctcccttttgtgYctccct -- 22 41634163 A ⇒ TA ⇒ T agagcRtaagccagagcRtaagcc --

한우 FABP4 유전자의 프라이머 염기서열Primer Sequence of Hanwoo FABP4 Gene 프라이머primer 서열order AFABP-인트론I-FAFABP-Intron I-F 5'-TTG TGC CTT GGG TGT TCT TT-3' (서열번호 1)5'-TTG TGC CTT GGG TGT TCT TT-3 '(SEQ ID NO: 1) AFABP-인트론I-RAFABP-Intron I-R 5'-TGG TTT GCT ATA GGC AGC AG-3' (서열번호 2)5'-TGG TTT GCT ATA GGC AGC AG-3 '(SEQ ID NO: 2) AFABP-인트론II-FAFABP-Intron II-F 5'-TGT TGT TTT TGG CAT TCA TTG-3' (서열번호 3)5'-TGT TGT TTT TGG CAT TCA TTG-3 '(SEQ ID NO: 3) AFABP-인트론II-RAFABP-Intron II-R 5'-TAC TGC TGG GGG CAC AGT AT-3' (서열번호 4)5'-TAC TGC TGG GGG CAC AGT AT-3 '(SEQ ID NO: 4) AFABP-엑손III-FAFABP-Exon III-F 5'-TTG TGC CTT GGG TGT TCT TT-3' (서열번호 5)5'-TTG TGC CTT GGG TGT TCT TT-3 '(SEQ ID NO: 5) AFABP-엑손III-RAFABP-Exon III-R 5'-TGG TTT GCT ATA GGC AGC AG-3' (서열번호 6)5'-TGG TTT GCT ATA GGC AGC AG-3 '(SEQ ID NO: 6)

게놈 DNA의 분리 추출은 MagExtractor Genomic DNA 정제 키트(Toyobo, Japan)를 이용하여 한우 583두의 혈액으로부터 수행하였다. 한우 지방세포-지방산 결합 단백질 유전자내의 각각 독립된 3좌위에 대한 PCR 반응을 위하여 2 ㎕의 PCR 반응 완충액(100 mM 트리스-HCl, pH 8.3; 500 mM KCl; 15 mM MgCl2), 2 ㎕의 2.5 mM dNTPs, 1 ㎕의 프라이머(10 pmole), 게놈 DNA 1 ㎕(100ng), 0.5 U의 Taq DNA 폴리머라아제(Promega, USA)를 넣고 최종 부피가 20 ㎕되게 조정하였다. PCR 수행은 PTC-225 tetrad PCR machine (MJ Research, Inc.)을 이용하여, 94℃에서 5분간 예비변성 후, 94℃에서 1분간 변성, 58℃에서 1분간 어닐링 및 72℃에서 2분간 확장하는 사이클을 35회 실시하였고, 마지막으로 72℃에서 10분간 더 반응시켰다. PCR이 끝난 후 5㎕의 PCR 증폭산물을 2%의 아가로오스 겔(agarose gel)을 이용하여 증폭여부를 확인하였다.Separation extraction of genomic DNA was performed from 583 blood of Hanwoo cattle using MagExtractor Genomic DNA Purification Kit (Toyobo, Japan). 2 μl of PCR reaction buffer (100 mM Tris-HCl, pH 8.3; 500 mM KCl; 15 mM MgCl 2 ), 2 μl of 2.5 mM for PCR reactions to independent tridents within the Hanwoo fat cell-fatty acid binding protein gene dNTPs, 1 μl of primer (10 pmole), 1 μl of genomic DNA (100 ng), 0.5 U of Taq DNA polymerase (Promega, USA) were added and the final volume was adjusted to 20 μl. PCR was performed using a PTC-225 tetrad PCR machine (MJ Research, Inc.), followed by premodification at 94 ° C for 5 minutes, denaturation at 94 ° C for 1 minute, annealing at 58 ° C for 1 minute, and expansion at 72 ° C for 2 minutes. The cycle was carried out 35 times and finally reacted at 72 ° C. for 10 minutes. After PCR, 5 μl of PCR amplification product was amplified using 2% agarose gel (agarose gel).

한우 개체별 유전자형 결정은 PCR-RFLP 방법을 이용하였는데, 개체별 PCR 증폭산물에서 5 ㎕씩을 각각 취하여 제한효소 Hpy188I(NEB), RsaI(Promega) 및 NlaIII(NEB)로 65℃ 및 37℃에서 각각 증폭산물을 완전히 절단 반응시켰다. 절단된 DNA 단편을 확인하기 위하여 2.5% 아가로오스 겔을 이용하여 전기영동을 실시하였고, EtBr 염색법으로 DNA 밴드를 검출하여 각 개체의 유전자형을 판정하였다.The genotyping was performed by PCR-RFLP method, and amplified by restriction enzymes Hpy188I (NEB), RsaI (Promega) and NlaIII (NEB) at 65 ° C and 37 ° C, respectively. The product was cleaved completely. To identify the cleaved DNA fragment, electrophoresis was performed using 2.5% agarose gel, and genotype of each individual was determined by detecting DNA bands by EtBr staining.

한우 개체별 유전자형의 판정은 도 1에서와 같이 인트론 1에 존재하는 C->G SNP의 경우 제한효소 Hpy188I에 의해 절단되어 328 bp 와 154 bp를 갖는 유전자형은 CC형으로 230 bp, 154 bp 및 98 bp를 갖는 유전자형은 GG형으로 그리고, 328 bp, 230 bp 및 154 bp를 갖는 유전자형은 GC형으로 판정하였다. 아울러 인트론 2의 A->T SNP 및 엑손 3의 A->G (Met->Val) SNP는 도 1과 같이 유전자형을 판정하였다.As for the genotype of Korean cattle, the genotypes of C-> G SNP present in intron 1 were cleaved by restriction enzyme Hpy188I as shown in FIG. 1, and 328 bp and 154 bp genotypes were 230 bp, 154 bp and 98. Genotypes with bp were determined to be GG type and genotypes with 328 bp, 230 bp and 154 bp were GC type. In addition, A-> T SNP of Intron 2 and A-> G (Met-> Val) SNP of Exon 3 were determined as shown in FIG. 1.

한우의 FABP4의 독립된 3개 SNP 좌위에 대한 유전자형에 따른 유전적 능력 차이를 규명하기 위해 혈통기록 및 환경요인 등을 고려한 아래와 같은 분석모형에 의한 유전능력 평가결과의 형질별 육종가(breeding value) 및 표현형 기록치를 이용하여 관련성 분석을 실시하였다.Breeding values and phenotypes of the traits of the results of the genetic ability evaluation based on the following analysis model considering the genealogy and environmental factors to determine the genetic differences of three independent SNP loci of FABP4 of Hanwoo Relevance analysis was performed using the records.

Figure 112006050071003-PAT00001
Figure 112006050071003-PAT00001

여기서,

Figure 112006050071003-PAT00002
: 표현형 능력기록,
Figure 112006050071003-PAT00003
: 전체 평균,
Figure 112006050071003-PAT00004
:
Figure 112006050071003-PAT00005
번째 연도-계절효과,
Figure 112006050071003-PAT00006
: 성별 효과,
Figure 112006050071003-PAT00007
: 도축일령에 대한 공변이, : 임의오차(random error)이다. here,
Figure 112006050071003-PAT00002
Phenotype record,
Figure 112006050071003-PAT00003
: Overall average,
Figure 112006050071003-PAT00004
:
Figure 112006050071003-PAT00005
Year-season effect,
Figure 112006050071003-PAT00006
: Gender effects,
Figure 112006050071003-PAT00007
: Covariation for slaughter age, This is a random error.

조사된 162두의 육종가 기록에 대해 FABP4 유전자형의 효과를 추정하기 위하여 SAS 8.1 Package/PC를 이용하여 아래와 같은 일반선형모형(Generalized Linear Model)을 적용한 최소제곱법(Least Squares Method)으로 통계 분석을 실시하였다. In order to estimate the effect of FABP4 genotype on the record of 162 breeders surveyed, statistical analysis was performed by using the least-squares method using the Generalized Linear Model using SAS 8.1 Package / PC. It was.

Figure 112006050071003-PAT00009
Figure 112006050071003-PAT00009

여기서,

Figure 112006050071003-PAT00010
: 육종가(breeding value),
Figure 112006050071003-PAT00011
: 전체평균,
Figure 112006050071003-PAT00012
: 유전자형 효과,
Figure 112006050071003-PAT00013
: 임의오차(random error)이다. here,
Figure 112006050071003-PAT00010
: Breeding value,
Figure 112006050071003-PAT00011
= Overall mean,
Figure 112006050071003-PAT00012
: Genotyping effect,
Figure 112006050071003-PAT00013
This is a random error.

한우 FABP4 유전자의 유전자형별 후대검정우 도체형질의 육종가 분석 결과Results of Breeding Value Analysis of Carcass Quality of Genotype Black Cows by Korean Genome of FABP4 SNPSNP 유전자형genotype 두수Head 도체중Conductor weight 배장근단면적Intestinal muscle area 등지방두께Back fat thickness 근내지방도Intramuscular fat map Hpy188I (C->G)Hpy188I (C-> G) CCCC 232232 1.9489±0.81631.9489 ± 0.8163 0.6481±0.22740.6481 ± 0.2274 -0.0032±0.0078-0.0032 ± 0.0078 0.0102±0.0409 a0.0102 ± 0.0409 a GCGC 288288 0.5135±0.73270.5135 ± 0.7327 0.5197±0.20410.5197 ± 0.2041 0.0084±0.00700.0084 ± 0.0070 0.0842±0.0368 ab0.0842 ± 0.0368 ab GGGG 5454 -0.5038±1.6920-0.5038 ± 1.6920 1.0182±0.47141.0182 ± 0.4714 0.0012±0.01610.0012 ± 0.0161 0.2334±0.0849 b0.2334 ± 0.0849 b RsaI (A->T)RsaI (A-> T) AAAA 242242 2.3678±0.79292.3678 ± 0.7929 0.6536±0.22260.6536 ± 0.2226 -0.0015±0.0076-0.0015 ± 0.0076 0.0024±0.0404 a 0.0024 ± 0.0404 a ATAT 274274 0.2476±0.75090.2476 ± 0.7509 0.5126±0.21090.5126 ± 0.2109 0.0082±0.00720.0082 ± 0.0072 0.1044±0.0383 ab 0.1044 ± 0.0383 ab TTTT 5656 -0.3939±1.6404-0.3939 ± 1.6404 1.0538±0.46061.0538 ± 0.4606 0.0141±0.01570.0141 ± 0.0157 0.2410±0.0837 b 0.2410 ± 0.0837 b Nla III (A->G)Nla III (A-> G) AAAA 5656 0.5996±1.66920.5996 ± 1.6692 0.8054±0.46410.8054 ± 0.4641 0.0258±0.01580.0258 ± 0.0158 -0.0348±0.0843 a -0.0348 ± 0.0843 a AGAG 250250 0.5398±0.79000.5398 ± 0.7900 0.3190±0.21960.3190 ± 0.2196 -0.0053±0.0075-0.0053 ± 0.0075 0.0158±0.0399 a 0.0158 ± 0.0399 a GGGG 270270 1.5362±0.76021.5362 ± 0.7602 0.8472±0.21130.8472 ± 0.2113 0.0064±0.00720.0064 ± 0.0072 0.1537±0.0384 b 0.1537 ± 0.0384 b

a, b : 다른 표기 사이에는 유의적인 차이가 인정됨(p<0.05).a, b: Significant difference is recognized between different notations (p <0.05).

한우 FABP4 유전자의 유전자형별 후대검정우 도체형질의 표현형 분석 결과Phenotypic Analysis of Genotypes of Subsequent Test Carcasses of Hanwoo FABP4 Gene SNPSNP 유전자형genotype 두수Head 도체중Conductor weight 배장근단면적Intestinal muscle area 등지방두께Back fat thickness 근내지방도Intramuscular fat map Hpy188I (C->G)Hpy188I (C-> G) CCCC 232232 311.737±0.669 a 311.737 ± 0.669 a 74.152±0.49174.152 ± 0.491 7.234±0.1967.234 ± 0.196 1.920±0.085 a1.920 ± 0.085 a GCGC 288288 313.778±0.592 ab 313.778 ± 0.592 ab 74.827±0.43574.827 ± 0.435 7.345±0.1737.345 ± 0.173 2.001±0.075 ab2.001 ± 0.075 ab GGGG 5454 315.069±1.304 b 315.069 ± 1.304 b 76.112±0.95876.112 ± 0.958 7.219±0.3827.219 ± 0.382 2.335±0.165 b2.335 ± 0.165 b RsaI (A->T)RsaI (A-> T) AAAA 242242 312.325±0.676 a 312.325 ± 0.676 a 74.070±0.49974.070 ± 0.499 7.288±0.1997.288 ± 0.199 1.882±0.087 a 1.882 ± 0.087 a ATAT 274274 314.191±0.599 b 314.191 ± 0.599 b 74.996±0.44274.996 ± 0.442 7.326±0.1767.326 ± 0.176 2.042±0.077 ab 2.042 ± 0.077 ab TTTT 5656 315.181±1.274 b 315.181 ± 1.274 b 76.040±0.94176.040 ± 0.941 7.399±0.3767.399 ± 0.376 2.309±0.163 b 2.309 ± 0.163 b NlaIII (A->G)NlaIII (A-> G) AAAA 5555 310.348±1.304 a 310.348 ± 1.304 a 74.841±0.96374.841 ± 0.963 7.733±0.3847.733 ± 0.384 1.866±0.167 a 1.866 ± 0.167 a AGAG 250250 313.186±0.631 b 313.186 ± 0.631 b 74.126±0.46674.126 ± 0.466 7.176±0.1867.176 ± 0.186 1.889±0.081 ab 1.889 ± 0.081 ab GGGG 270270 313.765±0.617 b 313.765 ± 0.617 b 75.194±0.45675.194 ± 0.456 7.314±0.1827.314 ± 0.182 2.161±0.079 b 2.161 ± 0.079 b

a, b : 다른 표기 사이에는 유의적인 차이가 인정됨(p<0.05).a, b: Significant difference is recognized between different notations (p <0.05).

통계분석 결과 표 3 및 표 4에서 보는 바와 같이 FABP4 유전자내의 독립된 3좌위와 육종가 및 표현형기록치와 관련성 분석하였다. 그 결과, 육종가에서 FABP4의 유전자형 중 Hpy188I 제한효소 절단 유전자형인 GG, RsaI의 TT 및 NlaIII의 GG형에서 근내지방도와 통계학적인 유의차를 얻었고, 특히, 표 4에서 보여지는 바와 같이 표현형기록치와 관련성 분석결과, Hpy188I (C⇒G), RsaI (A⇒T) 및 Nla III (A⇒G) 3좌위의 유전자형에서 동시에 도체중이 무거워지면서, 근내지방도가 우수해지는 효과가 있는 것으로 분석되었다(p<0.05). 그러므로 한우에 있어서 육량 및 육질 개량을 위한 우수 송아지 및 종축의 선발에 이 유전자형을 유전적 표지인자로 사용 가능함을 보여 준다.As a result of statistical analysis, as shown in Table 3 and Table 4, the correlation with the independent three positions, breeders and phenotypic records in FABP4 gene was analyzed. As a result, the intramuscular fat and statistically significant differences were obtained between the Hpy188I restriction enzyme cleavage genotypes GG, TT of RsaI, and GG of NlaIII among the genotypes of FABP4. As a result, it was analyzed that the genotypes of Hpy188I (C⇒G), RsaI (A⇒T), and Nla III (A⇒G) tridents resulted in heavy carcass weight and good intramuscular fat (p < 0.05). Therefore, this genotype can be used as a genetic marker for the selection of excellent calf and breeders for meat and meat improvement in Korean cattle.

상기에서 살펴본 바와 같이, 본 발명에서 밝혀진 한우의 도체중과 근내지방도에 유의적인 효과를 가지는 FABP4 (Adipoctye-Fatty Acid Binding Protein) 유전자의 독립된 3좌위인 Hpy188I (C⇒G), RsaI (A⇒T) 및 NlaIII (A⇒G)의 유전자형을 이용하여 한우 개체별 유전적 자질을 판정함으로써 비육 밑소의 조기 선발, 육종가의 정확한 추정 등에 활용이 가능하다.As described above, Hpy188I (C⇒G), RsaI (A⇒T), which are independent three-positions of FABP4 (Adipoctye-Fatty Acid Binding Protein) gene, have a significant effect on carcass weight and intramuscular fat of Hanwoo, as disclosed in the present invention. ) And NlaIII (A⇒G) genotypes to determine the genetic characteristics of each individual cattle, it can be used for early selection of fattening cattle, accurate estimation of breeding value.

<110> REPUBLIC OF KOREA(MANAGEMENT : RURAL DEVELOPMENT ADMINISTRATION) <120> enetic marker of adipocyte-fatty acid binding protein gene related the intramuscular fat content(marbling) in beef cattle <160> 11 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron I forward primer <400> 1 ttgtgccttg ggtgttcttt 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron I reverse primer <400> 2 tggtttgcta taggcagcag 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron II forward primer <400> 3 tgttgttttt ggcattcatt g 21 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron II reverse primer <400> 4 tactgctggg ggcacagtat 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-exon III forward primer <400> 5 ttgtgccttg ggtgttcttt 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-exon III reverse primer <400> 6 tggtttgcta taggcagcag 20 <210> 7 <211> 4236 <212> DNA <213> Bovine FABP4 gene <220> <221> CDS <222> (1)..(72) <223> exon 1 of bovine FABP4 gene <220> <221> CDS <222> (2783)..(2956) <223> exon 2 of bovine FABP4 gene <220> <221> CDS <222> (3551)..(3655) <223> exon 3 of bovine FABP4 gene <220> <221> CDS <222> (4116)..(4163) <223> exon 4 of bovine FABP4 gene <400> 7 atg tgt gat gca ttt gta ggt acc tgg aaa ctt gtc tcc agt gaa aac 48 Met Cys Asp Ala Phe Val Gly Thr Trp Lys Leu Val Ser Ser Glu Asn 1 5 10 15 ttt gat gat tac atg aaa gaa gtg ggtaagga aatgcattgt tgaatggctg 100 Phe Asp Asp Tyr Met Lys Glu Val 20 ggcttataac tttttctcta ggaaagagag gcctatggtt ctttttcact ctggaagcat 160 tggtctgaca tgaaatgctg ctttctatac agaggtgata aaaataacaa gtgagaatca 220 tttgtttcca gaaacattct agatggaagt cacaactaat acttcctaaa ggtgaactga 280 tatgaaaaag agtataagtg gcattgtata gaaatgacaa gattttattt gattgctaaa 340 cttgagtatc tttggtttac ttggctttct gcctcttttt cattcattcc ttttgcatat 400 aaaataagca ataacttgga aataagccta tatttattcc ctagcatgtg ataatttcca 460 tttgaaactg aaagcactgt atttatcatc ttaacataat tttttgagta atgtatgtta 520 accaattatt ttattgtttt aaagtcaatc tgttaaaatg tgtggttgta ttatggaagg 580 aaaaaatcta atttctgaaa atgtttcctt raaagtctta gatatttgtt tgrcttaata 640 ttacaaggca tgacacagaa ttattgagaa tagaggcttt caagtttttg gattctgcta 700 agactgcctg tatgttcccc agataattac aaaggtaaag caaaggagca gagaggcggg 760 aattcagtca gacagcccta cccatgcatg atgaagatgt atccatgggt tgcaaattaa 820 ggagggattt ttttcacaat atctttatta gaggttataa tttaatccct aaatctcaag 880 acttaacctt cttctaaatc ttatagttat ttcttgcttg tttataacta ctgttttcca 940 tgtttataga caagaaggaa atcaggaaca tcaatatgag tgagaactag ttttccagac 1000 ttytgactag ttgtaactca ataaactaat cagtatgcac tgatattgat attayaggac 1060 ttgagtctat ttacctattt atttttagaa gcattttaca cttttagtat tttatttaaa 1120 ttcttatacc attgcgtttg ggcttcctat rtggtgctag tggtaaagaa cccacctgcc 1180 aattcaggag atttaagaga cagagggttc gattcctggg tcaggaagat tcccctagag 1240 gagggcatgg caacctactt cagtattctt gcctggagaa tcccatgaac agaggagcct 1300 ggtgcactat ggttcatagg gttgcaaaga gtcagacatg actgaagaga cttagcacac 1360 atgcaggcac cacagtcttt taaaatgatt aagtattgat gataatgaaa gacaataaga 1420 tacattctga aattagctgc tctatttgat taccttcagt cagcaggtat cattttcatc 1480 agtcaaatga aaacagaatc aagcaagagg agagtgtgat gttcctaagc ttggagaatc 1540 tggaccacac ccgatgtctc atgcactcac tgtgctgact cactccatga atttgaggtt 1600 acattttgtt aacagagaca agacttgatg aagcagcata ttatcttccc acctgtcacc 1660 actttcagac agggggctaa atcagagcct tagcattgct tgttttgtat gatatatcgc 1720 ctccccacta ttcttaaatt ttggtatcta tacatagtca aagaagtaga tttgtacatg 1780 tgcatttaaa aagtccaaca cacctatgac tgtcataaat ctctataaaa catgcagtga 1840 tggtaaggta ctgttttcta tgttaagttg taaataattc aacccaaagc tgtaacatcc 1900 cccaaaacac attcatattt tttctttttt taaaaaagag gagtttgcca cttaagtgat 1960 tgggaagcaa agtattaatt tctgaactta ctaaaataac gatttcttaa actttgaaat 2020 tttcatttga tgcctaagcc ctctttaaat ttttctataa tttattggga ataccatcac 2080 cattctaaaa tggtatgaat gcaagaaaag ttttgccttt cagaaaaagt aatcaagatc 2140 ttcatttcag agtaattatg aaaagccaag aaaactaggt gtggcagcga atcccattaa 2200 atgtggkttt tgtacaccca aattaaattt taactgagtg actctctgtt ggtyycctga 2260 atcctattaa aagttctact tttgactatg gattagatca tttttgttat tcaagaagca 2320 atggttgaga ttcccaagta tttcatgaac tgagtcaaat gaagctggct gctctcatgg 2380 ttaagatgga cagttatttt gggagtgcag tgatttttac gatatgtatg tcccagaaaa 2440 tttaaccttt tacatatgct tatagaaaat taacaatggg tctcctgaag agtctcatct 2500 gaaacccaga gggggaaatt taaagaacag acttaatatc taattgtgcc ttgggtgttc 2560 tttaagtatt agctgatttt acttattttc aggagatgtt taaataaaga ctttaataat 2620 aaactcactg gagtatttct cctttaatat acacactgga ctgtagatag gtatatgggc 2680 acacatgcgt ctgtctaaat acacacacac acacctgctc tttcttagat aaatatatga 2740 gagaaaaact gtatacttga catttttctt ttccsaacta ta ggc gtg ggc 2791 Gly Val Gly 1 ttt gct acc agg aaa gtg gct ggc atg gcc aaa ccc act ttg atc atc 2839 Phe Ala Thr Arg Lys Val Ala Gly Met Ala Lys Pro Thr Leu Ile Ile 5 10 15 agt ttg aat ggg ggt gtg gtc acc att aaa tca gaa agc acc ttt aaa 2887 Ser Leu Asn Gly Gly Val Val Thr Ile Lys Ser Glu Ser Thr Phe Lys 20 25 30 35 aat act gag att tcc ttc aaa ttg ggc cag gaa ttt gat gaa rtc act 2935 Asn Thr Glu Ile Ser Phe Lys Leu Gly Gln Glu Phe Asp Glu Xaa Thr 40 45 50 cca gat gac agg aaa gtc aag gtga gaaataaaga actggagcag agtaaaaaac 2990 Pro Asp Asp Arg Lys Val Lys 55 ctgatttata aatgactgct gcctatagca aaccattttg tagaaggagg aaagccattc 3050 cattataagc caaaaatctc agattgctag atctgaacca tgttaccttt gatatttagc 3110 tggtgaattt tctcccattt aataaaattg tccttattac tttaaaaatg tttaacataa 3170 taatttactt gtcataccat atatatgtgt gtatttatat atatatatat atatattatt 3230 tgaagtaaat tgaagtaaca taacaatgtt agagaacttt taaaagagtg gggggaaagg 3290 aaaaaaaaaa cccctatgat gctattccac ataaatttat tatctatatt ctttcacagt 3350 attttttttt caaatgcatg tttgtataat attctgatca taatatacat gtaattttgt 3410 atgttgtttt tggcattcat tgttttmttt tgcaacattt tcttgtaatt tagaattgct 3470 aagwacctca aaataagcaa ataaaagcac tctatttttt ttccctccat cattgtaatc 3530 acttttaatt atccccacag agc atc gta aac tta gat gaa ggt gct ctg gta 3583 Ser Ile Val Asn Leu Asp Glu Gly Ala Leu Val 1 5 10 caa gta caa aac tgg gat gga aaa tca acc acc ata aag aga aaa ctc 3631 Gln Val Gln Asn Trp Asp Gly Lys Ser Thr Thr Ile Lys Arg Lys Leu 15 20 25 rtg gat gat aag atg gtg cts gtg agtat cttctcacta cttaattcta 3680 Xaa Asp Asp Lys Met Val Xaa Val 30 35 gatttyagtg ctaggtcatc ccataatygt tatcctacct agagaaatag acaatcgccc 3740 ttgtagaatg aaaagttagt ctattgggat tatggtttca ctctgacaat tatccttcta 3800 agctccgtct aggtatactg tgcccccagc agtattttct tatccctctc aatgtgaacc 3860 gtattgtatt gtgcatttct aattatgttt ttcactcacc acatagatgg taagattcct 3920 tgaggccaag tcttgtatct tcttgatctt tgtgyctccc tagtttatta caatatcagg 3980 tatataagaa gagccaagag ggaatatctt ttgatgaaca ttttttcctg ctcaacattg 4040 aaggagacaa taaataaata aaacataagt tgtttagtcc tgaggatttt accaatattt 4100 tgcttttgtg cctag gaa tgt gtc atg aat ggt gtc act gcc acc aga 4148 Glu Cys Val Met Asn Gly Val Thr Ala Thr Arg 1 5 10 gtt tat gag aga gcr taagcca agggatattg aaatggatga cgtttgcatc 4200 Val Tyr Glu Arg Xaa 15 gaactccatg actttctgct ggatacgttg tccaaa 4236 <210> 8 <211> 24 <212> PRT <213> Bovine FABP4 gene <400> 8 Met Cys Asp Ala Phe Val Gly Thr Trp Lys Leu Val Ser Ser Glu Asn 1 5 10 15 Phe Asp Asp Tyr Met Lys Glu Val 20 <210> 9 <211> 58 <212> PRT <213> Bovine FABP4 gene <400> 9 Gly Val Gly Phe Ala Thr Arg Lys Val Ala Gly Met Ala Lys Pro Thr 1 5 10 15 Leu Ile Ile Ser Leu Asn Gly Gly Val Val Thr Ile Lys Ser Glu Ser 20 25 30 Thr Phe Lys Asn Thr Glu Ile Ser Phe Lys Leu Gly Gln Glu Phe Asp 35 40 45 Glu Xaa Thr Pro Asp Asp Arg Lys Val Lys 50 55 <210> 10 <211> 35 <212> PRT <213> Bovine FABP4 gene <400> 10 Ser Ile Val Asn Leu Asp Glu Gly Ala Leu Val Gln Val Gln Asn Trp 1 5 10 15 Asp Gly Lys Ser Thr Thr Ile Lys Arg Lys Leu Xaa Asp Asp Lys Met 20 25 30 Val Xaa Val 35 <210> 11 <211> 16 <212> PRT <213> Bovine FABP4 gene <400> 11 Glu Cys Val Met Asn Gly Val Thr Ala Thr Arg Val Tyr Glu Arg Xaa 1 5 10 15 <110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> enetic marker of adipocyte-fatty acid binding protein gene          related the intramuscular fat content (marbling) in beef cattle <160> 11 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron I forward primer <400> 1 ttgtgccttg ggtgttcttt 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron I reverse primer <400> 2 tggtttgcta taggcagcag 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron II forward primer <400> 3 tgttgttttt ggcattcatt g 21 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-intron II reverse primer <400> 4 tactgctggg ggcacagtat 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-exon III forward primer <400> 5 ttgtgccttg ggtgttcttt 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> AFABP-exon III reverse primer <400> 6 tggtttgcta taggcagcag 20 <210> 7 <211> 4236 <212> DNA <213> Bovine FABP4 gene <220> <221> CDS (222) (1) .. (72) <223> exon 1 of bovine FABP4 gene <220> <221> CDS (222) (2783) .. (2956) <223> exon 2 of bovine FABP4 gene <220> <221> CDS <222> (3551) .. (3655) <223> exon 3 of bovine FABP4 gene <220> <221> CDS (4116) .. (4163) <223> exon 4 of bovine FABP4 gene <400> 7 atg tgt gat gca ttt gta ggt acc tgg aaa ctt gtc tcc agt gaa aac 48 Met Cys Asp Ala Phe Val Gly Thr Trp Lys Leu Val Ser Ser Glu Asn   1 5 10 15 ttt gat gat tac atg aaa gaa gtg ggtaagga aatgcattgt tgaatggctg 100 Phe Asp Asp Tyr Met Lys Glu Val              20 ggcttataac tttttctcta ggaaagagag gcctatggtt ctttttcact ctggaagcat 160 tggtctgaca tgaaatgctg ctttctatac agaggtgata aaaataacaa gtgagaatca 220 tttgtttcca gaaacattct agatggaagt cacaactaat acttcctaaa ggtgaactga 280 tatgaaaaag agtataagtg gcattgtata gaaatgacaa gattttattt gattgctaaa 340 cttgagtatc tttggtttac ttggctttct gcctcttttt cattcattcc ttttgcatat 400 aaaataagca ataacttgga aataagccta tatttattcc ctagcatgtg ataatttcca 460 tttgaaactg aaagcactgt atttatcatc ttaacataat tttttgagta atgtatgtta 520 accaattatt ttattgtttt aaagtcaatc tgttaaaatg tgtggttgta ttatggaagg 580 aaaaaatcta atttctgaaa atgtttcctt raaagtctta gatatttgtt tgrcttaata 640 ttacaaggca tgacacagaa ttattgagaa tagaggcttt caagtttttg gattctgcta 700 agactgcctg tatgttcccc agataattac aaaggtaaag caaaggagca gagaggcggg 760 aattcagtca gacagcccta cccatgcatg atgaagatgt atccatgggt tgcaaattaa 820 ggagggattt ttttcacaat atctttatta gaggttataa tttaatccct aaatctcaag 880 acttaacctt cttctaaatc ttatagttat ttcttgcttg tttataacta ctgttttcca 940 tgtttataga caagaaggaa atcaggaaca tcaatatgag tgagaactag ttttccagac 1000 ttytgactag ttgtaactca ataaactaat cagtatgcac tgatattgat attayaggac 1060 ttgagtctat ttacctattt atttttagaa gcattttaca cttttagtat tttatttaaa 1120 ttcttatacc attgcgtttg ggcttcctat rtggtgctag tggtaaagaa cccacctgcc 1180 aattcaggag atttaagaga cagagggttc gattcctggg tcaggaagat tcccctagag 1240 gagggcatgg caacctactt cagtattctt gcctggagaa tcccatgaac agaggagcct 1300 ggtgcactat ggttcatagg gttgcaaaga gtcagacatg actgaagaga cttagcacac 1360 atgcaggcac cacagtcttt taaaatgatt aagtattgat gataatgaaa gacaataaga 1420 tacattctga aattagctgc tctatttgat taccttcagt cagcaggtat cattttcatc 1480 agtcaaatga aaacagaatc aagcaagagg agagtgtgat gttcctaagc ttggagaatc 1540 tggaccacac ccgatgtctc atgcactcac tgtgctgact cactccatga atttgaggtt 1600 acattttgtt aacagagaca agacttgatg aagcagcata ttatcttccc acctgtcacc 1660 actttcagac agggggctaa atcagagcct tagcattgct tgttttgtat gatatatcgc 1720 ctccccacta ttcttaaatt ttggtatcta tacatagtca aagaagtaga tttgtacatg 1780 tgcatttaaa aagtccaaca cacctatgac tgtcataaat ctctataaaa catgcagtga 1840 tggtaaggta ctgttttcta tgttaagttg taaataattc aacccaaagc tgtaacatcc 1900 cccaaaacac attcatattt tttctttttt taaaaaagag gagtttgcca cttaagtgat 1960 tgggaagcaa agtattaatt tctgaactta ctaaaataac gatttcttaa actttgaaat 2020 tttcatttga tgcctaagcc ctctttaaat ttttctataa tttattggga ataccatcac 2080 cattctaaaa tggtatgaat gcaagaaaag ttttgccttt cagaaaaagt aatcaagatc 2140 ttcatttcag agtaattatg aaaagccaag aaaactaggt gtggcagcga atcccattaa 2200 atgtggkttt tgtacaccca aattaaattt taactgagtg actctctgtt ggtyycctga 2260 atcctattaa aagttctact tttgactatg gattagatca tttttgttat tcaagaagca 2320 atggttgaga ttcccaagta tttcatgaac tgagtcaaat gaagctggct gctctcatgg 2380 ttaagatgga cagttatttt gggagtgcag tgatttttac gatatgtatg tcccagaaaa 2440 tttaaccttt tacatatgct tatagaaaat taacaatggg tctcctgaag agtctcatct 2500 gaaacccaga gggggaaatt taaagaacag acttaatatc taattgtgcc ttgggtgttc 2560 tttaagtatt agctgatttt acttattttc aggagatgtt taaataaaga ctttaataat 2620 aaactcactg gagtatttct cctttaatat acacactgga ctgtagatag gtatatgggc 2680 acacatgcgt ctgtctaaat acacacacac acacctgctc tttcttagat aaatatatga 2740 gagaaaaact gtatacttga catttttctt ttccsaacta ta ggc gtg ggc 2791                                                        Gly val gly                                                          One ttt gct acc agg aaa gtg gct ggc atg gcc aaa ccc act ttg atc atc 2839 Phe Ala Thr Arg Lys Val Ala Gly Met Ala Lys Pro Thr Leu Ile Ile       5 10 15 agt ttg aat ggg ggt gtg gtc acc att aaa tca gaa agc acc ttt aaa 2887 Ser Leu Asn Gly Gly Val Val Thr Ile Lys Ser Glu Ser Thr Phe Lys  20 25 30 35 aat act gag att tcc ttc aaa ttg ggc cag gaa ttt gat gaa rtc act 2935 Asn Thr Glu Ile Ser Phe Lys Leu Gly Gln Glu Phe Asp Glu Xaa Thr                  40 45 50 cca gat gac agg aaa gtc aag gtga gaaataaaga actggagcag agtaaaaaac 2990 Pro Asp Asp Arg Lys Val Lys              55 ctgatttata aatgactgct gcctatagca aaccattttg tagaaggagg aaagccattc 3050 cattataagc caaaaatctc agattgctag atctgaacca tgttaccttt gatatttagc 3110 tggtgaattt tctcccattt aataaaattg tccttattac tttaaaaatg tttaacataa 3170 taatttactt gtcataccat atatatgtgt gtatttatat atatatatat atatattatt 3230 tgaagtaaat tgaagtaaca taacaatgtt agagaacttt taaaagagtg gggggaaagg 3290 Aaaaaaaaaaa cccctatgat gctattccac ataaatttat tatctatatt ctttcacagt 3350 attttttttt caaatgcatg tttgtataat attctgatca taatatacat gtaattttgt 3410 atgttgtttt tggcattcat tgttttmttt tgcaacattt tcttgtaatt tagaattgct 3470 aagwacctca aaataagcaa ataaaagcac tctatttttt ttccctccat cattgtaatc 3530 acttttaatt atccccacag agc atc gta aac tta gat gaa ggt gct ctg gta 3583                       Ser Ile Val Asn Leu Asp Glu Gly Ala Leu Val                         1 5 10 caa gta caa aac tgg gat gga aaa tca acc acc ata aag aga aaa ctc 3631 Gln Val Gln Asn Trp Asp Gly Lys Ser Thr Thr Ile Lys Arg Lys Leu              15 20 25 rtg gat gat aag atg gtg cts gtg agtat cttctcacta cttaattcta 3680 Xaa Asp Asp Lys Met Val Xaa Val          30 35 gatttyagtg ctaggtcatc ccataatygt tatcctacct agagaaatag acaatcgccc 3740 ttgtagaatg aaaagttagt ctattgggat tatggtttca ctctgacaat tatccttcta 3800 agctccgtct aggtatactg tgcccccagc agtattttct tatccctctc aatgtgaacc 3860 gtattgtatt gtgcatttct aattatgttt ttcactcacc acatagatgg taagattcct 3920 tgaggccaag tcttgtatct tcttgatctt tgtgyctccc tagtttatta caatatcagg 3980 tatataagaa gagccaagag ggaatatctt ttgatgaaca ttttttcctg ctcaacattg 4040 aaggagacaa taaataaata aaacataagt tgtttagtcc tgaggatttt accaatattt 4100 tgcttttgtg cctag gaa tgt gtc atg aat ggt gtc act gcc acc aga 4148                       Glu Cys Val Met Asn Gly Val Thr Ala Thr Arg                         1 5 10 gtt tat gag aga gcr taagcca agggatattg aaatggatga cgtttgcatc 4200 Val Tyr Glu Arg Xaa              15 gaactccatg actttctgct ggatacgttg tccaaa 4236 <210> 8 <211> 24 <212> PRT <213> Bovine FABP4 gene <400> 8 Met Cys Asp Ala Phe Val Gly Thr Trp Lys Leu Val Ser Ser Glu Asn   1 5 10 15 Phe Asp Asp Tyr Met Lys Glu Val              20 <210> 9 <211> 58 <212> PRT <213> Bovine FABP4 gene <400> 9 Gly Val Gly Phe Ala Thr Arg Lys Val Ala Gly Met Ala Lys Pro Thr   1 5 10 15 Leu Ile Ile Ser Leu Asn Gly Gly Val Val Thr Ile Lys Ser Glu Ser              20 25 30 Thr Phe Lys Asn Thr Glu Ile Ser Phe Lys Leu Gly Gln Glu Phe Asp          35 40 45 Glu Xaa Thr Pro Asp Asp Arg Lys Val Lys      50 55 <210> 10 <211> 35 <212> PRT <213> Bovine FABP4 gene <400> 10 Ser Ile Val Asn Leu Asp Glu Gly Ala Leu Val Gln Val Gln Asn Trp   1 5 10 15 Asp Gly Lys Ser Thr Thr Ile Lys Arg Lys Leu Xaa Asp Asp Lys Met              20 25 30 Val Xaa Val          35 <210> 11 <211> 16 <212> PRT <213> Bovine FABP4 gene <400> 11 Glu Cys Val Met Asn Gly Val Thr Ala Thr Arg Val Tyr Glu Arg Xaa   1 5 10 15

Claims (3)

1) 서열번호 7의 한우 FABP4 유전자로부터 발견되고 도 1에 도시된 17개의 SNP 좌위 판독을 이용하여 연관군의 유전자형을 판별하는 단계;1) determining the genotype of the associated group using the 17 SNP locus reads found from the Hanwoo FABP4 gene of SEQ ID NO: 7 and shown in FIG. 1; 2) 서열번호 1 내지 서열번호 6의 PCR 프라이머를 이용하여 서열번호 7의 한우 FABP4 유전자를 주형으로 94℃에서 5분간 예비변성 후, 94℃에서 1분간 변성, 58℃에서 1분간 어닐링 및 72℃에서 2분간 확장하는 사이클을 35회 실시하고, 마지막으로 72℃에서 10분간 더 반응시켜 PCR 증폭하는 단계;2) After preliminary modification of the Hanwoo FABP4 gene of SEQ ID NO: 7 as a template for 5 minutes at 94 ° C using PCR primers of SEQ ID NO: 1 to SEQ ID NO: 6, denaturation at 94 ° C for 1 minute, annealing at 58 ° C for 1 minute, and 72 ° C. 35 cycles of expansion for 2 minutes at and finally PCR amplification by further reacting at 72 ° C. for 10 minutes; 3) 상기 증폭산물을 제한효소 Hpy188I, RsaI 및 NlaIII로 절단하여 전기영동을 실시하는 단계; 및3) electrophoresis by cleaving the amplified product with restriction enzymes Hpy188I, RsaI and NlaIII; And 4) 상기 전기영동 결과물에 대한 사진 및 이미지를 확보하여 개체별 유전자형을 결정하는 단계;4) determining a genotype for each individual by obtaining a photograph and an image of the electrophoretic result; 를 포함하는 유전적 자질을 판단하기 위한 DNA 표지인자를 선발하는 방법.Method for selecting a DNA marker for determining the genetic qualities comprising. 제 1항에 있어서, 상기 4) 단계에서 제한효소 절편길이 다양성(Restriction fragment length polymorphism, RFLP) 분석, 이형접합자 분석(heteroduplex analysis), 단일 나선의 구조적 다형성(single strand conformational polymorphism, SSCP) 및 테크맨 프로브법(Taqman probe method) 중 하나 이상의 과정을 더 거침으로써 개체별 유전자형을 결정함을 특징으로 하는 방법.According to claim 1, Restriction fragment length polymorphism (RFLP) analysis, heteroduplex analysis, single strand conformational polymorphism (SSCP) and Techman in step 4) A method characterized by determining the individual genotype by further undergoing one or more of the Taqman probe methods. 제 1항 또는 제 2항에 의한 방법으로 선발되고,Selected by the method according to claim 1, 지방세포(adipocyte)에서 발현되는 지방산 결합 단백질(fatty acid binding protein, FABP4) 유전자의 변이 부위 중 도 1에 도시된 Hpy188I (C⇒G), RsaI (A⇒T) 및 NlaIII (A⇒G) 3좌위를 인지하도록 제작된 DNA 표지인자.Among the mutation sites of the fatty acid binding protein (FABP4) gene expressed in adipocytes, Hpy188I (C⇒G), RsaI (A⇒T) and NlaIII (A⇒G) 3 shown in FIG. DNA markers designed to recognize loci.
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