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소 성장호르몬 유전자의 Exon 5번에서의 새로운 다형성 연구
윤두학,김태헌,이경희,박응우,이학교,정일정,홍기창 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.1
성장호르몬 유전자는 하나의 작은 공통 선조 유전자로부터 아주 오랜 기간동안 유전자 중복에 의해 진화되어 온 그룹들 중의 하나이다. 이들에 속하는 유전자들은 동물 종간에 구조적인 상동성과 기능적 공통성 등 유사성이 비교적 높게 나타난다. 이런 연구결과들을 근거로 하여 소 성장호르몬 유전자에서 아미노산을 암호화하는 영역으로부터 새로운 아미노산의 변이(missense mutation)를 검출하였고, 이 변이의 대립유전자 빈도는 소(cattle)의 종(species) 및 품종의 지리적 분포에 따라 일정한 경향 치를 보여 주었다. 한편 변경되어진 아미노산은 Tryptophan으로 이는 생물체에 존재하는 많은 단백질들을 구성하는 아미노산중에서도 그 출현빈도가 가장 낮은 것이다. 또한 검출된 변이는 성장호르몬이 그의 수용체와 강하게 결합하는 부위로서, 성장호르몬의 구조적 변이를 초래하여 수용체와의 결합이 비정상적으로 이루어져, 이후 성장호르몬이 표적세포로의 신호전달과 같은 역할을 제대로 수행치 못하게 되고, 이로 인하여 가축의 표현되어지는 경제형질에 영향을 미칠 것으로 추정된다. 그러므로 이러한 대립유전자를 보유하는 개체는 집단에서 제거하는 방법에 의한 개량이 가능할 것으로 사료된다. Growth Hormone (GH) gene is a member of gene family through the evolutionary process from a small common ancestral gene by a series of gene duplications. The role of the GH in growth and performance controls has bees extensively studied in human, mice and livestock. Many researchers have considered GH as a strong candidate gene for evaluation of genetic polymorphisms that could be associated with economic traits in cattle. We report here a novel missense mutation within the exon 5 of the bovine Growth Hormone (bGH) gene. We could amplified 522 bp fragments from eight unrelated Hanwoo cattle by PCR, the, subsequently cloned and sequenced. An Msp I RFLP corresponding to a C to T transition was observed at position 2258 nt. From this result, we could predict a missense mutation (Arg to Trp) at codon 166 in a highly conserved region among many mammals. Codominant Mendelian segregation of the two alleles, Msp I (+) and Msp I (-), was observed in two full-sib F2 families (n = 32, African taurine Bos taurus × African zebu Bos indicus) and eight half-sib Hanwoo families. For the availability of genetic marker, we have performed PCR-RFLP with a large number of individual animals from 15 different cattle breeds (European and Asian taurines. and African indicines). Consideration of breed frequencies of Msp I (=) allele in relation to breed type and their geographic origins, shows higher frequencies in humped breeds or Asian cattle breeds than in humpless or European breeds. This result indicates that the missense mutation can be contributed the functional significance such as the signal transduction through the receptor binding also may be used as a marker for selection of the economic traits in Hanwoo.
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돼지 염색체 6번의 연관지도 및 양적형질 유전자좌위 탐색
이혜영,최봉환,김태헌,박응우,윤두학,이학교,전광주,정일정,홍기창 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.6
The objective of this study was to identify the quantitative traits loci(QTL) for economically important traits such as growth, carcass and meat quality on pig chromosome 6. A three generation resource population was constructed from cross between Korean native boars and Landrace sows. A total of 240 F_(2) animals were produced using intercross between 10 boars and 31 sows of F_(1) animals. Phenotypic data including body weight at 3 weeks, backfat thickness, muscle ?, shear force and crude protein level were collected from F_(2) animals. Animals including grandparents(F_(0)), parents(F_(1)) and offspring(F_(2) were genotyped for 29 microsatellite markers and PCR-RFLP marker on chromosome 6. The linkage analysis was performed using CRI-MAP software version 2.4(Green et al., 1990) with FIXED option to obtain the map distances. The total length of SSC6 linkage map estimated in this study was 169.3cM. The average distance between adjacent markers was 6.05cM. For mapping of QTL, we used F_(2) QTL. Analysis Servelt of QTL express, a web-based QTL mapping tool(http://qtl.cap.ed.ac.uk). Five QTLs were detected at 5% chromosome-wide level for body weight of 3 weeks of age, shear force meat ? at 24 hours after slaughtering, backfat thickness and crude protein level on SSC6.
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한우 경제형질에 미치는 Mitochondrial DNA D-loop 영역의 염기서열 변이효과
오재돈,윤두학,공홍식,임현진,이학교,조병욱,홍기창,전광주 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.6
This study was performed to analyse the sequences of variations of mtDNA D-loop and their effects on carcass traits in Hnawoo(Korean cattle). The resulting sequences were compared with perviously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty five polymorphic sites by nucleotide substitution were found in mtDNA of Hanwoo. The frequencies of positions at 169, 16042, 16093. 16119, 16255 and 16302 nt with high levels of sequence polymorphism were 0.891, 0.117, 0.109, 0.182, 0.197 and 0.117, respectively. The substitution effect at 169 and 16119 nt was found significant on marbling score. Also substitution effect at 169 and 16042 nt was highly significant(p<0.01) on backfat, thickness. Polymorphisrn of mtDNA sequence in D-loop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.
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재래돼지 미토콘드리아 게놈내 D-loop 영역의 염기서열 분석
정일정,김태헌,윤두학,조진기,이효신 한국축산학회 1997 한국축산학회지 Vol.39 No.3
The objective of this study was to investigate differences in mtDNA D-loop sequence between Korean native pig and other breeds. When digested with BamH I/Cla I, D-loop region was found in 1.03 kb and 1.59 kb fragments. Both fragments containing D-loop region were subcloned into plasmid vector of pBluescript II SK^+ and sequenced. After sequencing 2616 by in this study, D-loop region was 1313 by and the rest of them were genes for cytochrome b, 12S rRNA, tRNA^(Pro) tRNA^(Phe) and tRNA^(Thr). The twenty-eight tandem repeats of 10bp, CGTGCGTACA, were found within D-loop region of mtDNA in Korean native pig. The sequence variation between Korean native pig and Meishan pig was observed for only 3 nucleotides substitution and 2 nucleotides deletion. However, the sequence difference between the Korean native pig and European pig showed 16 nucleotide substitution, 1 nucleotide deletion and 1 nucleotide insertion. The highly variable region among the three breeds was located between tRNA^(Pro) gene and CSB 1.
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재래돼지 Melanocortin Receptor 1(MCIR)의 변이에 관한 연구
한재용,정일정,김태헌,윤두학,연성흠,최봉환,노환국,김남순,이화진 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.3
The melanocortin 1 receptor (MC1R) gene is the major candidate gene for coat color in pig since it is assumed to be controlled by an allele at the Extension (E) locus. A 702 by fragments of the MC1R gene were amplified by PCR, cloned and sequenced from 6 heads of Korean native pig. Of them, 4 pigs possessed allele#2 which is associated with black coat color in pig, the others had allele#1 which revealed a unique allele in European wild boar reported by Kijas et al. (1998). PCR-RFLP was carried out to observe their alleles from 6 different pig breeds including Korean native pig, Berkshire, Duroc, Landrace, Large White, and Meishan pig. Two pairs of primers were designed for amplifying the MC1R gene fragment corresponding to five polymorphic sites. PCR products were digested 5 different restriction enzymes. Nsp I , MspA 1 I , Rca I , Hha I and Acc II , and electrophoresed in 2.5% Metaphor agarose gel. All breeds except for Korean native and Meishan breeds were the same alleles reported by Kijas et al. (1998). Of 9 Meishan pigs tested in this study, 4 heads possessed the same allele, 5 heads were different alleles reported by Kijas et al. (1998). A total of 15 different alleles revealed in Korean native pig. The results of this study strongly suggest that black coat color in Korean native breed was associated with a constitutively active receptor at codon 99 (L99P).
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소 품종별 Melanocortin Receptor 1(MC1R) 유전자의 유전자형 빈도에 관한 연구
한재용,김경남,오성종,정일정,김태헌,윤두학,박응우,이혜영,탁태영 한국동물자원과학회 2000 한국축산학회지 Vol.42 No.6
The melanocortin 1 receptor (MC1R) plays a central role in regulation of eumelanin(black/brown) and phaeomelanin (red / yellow) synthesis within the mammalian melanocyte and encoded by the classical Extension (E) coat color locus. The objectives of this study were carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no. Y19103). A size of 350 by was amplified by polymerase chain reaction(PCR), digested with each or together of two different restriction enzyme, BsrFI and MspAII, and electrophoresed in 2.5% or 4% Metaphore agarose gel for determination of genotypes. One thousand and forty four samples including eight different cattle breeds and imported beef were determinated their genotypes by PCR-RFLP. Genotype frequencies of Hanwoo were 0.10 in E e and 0.90 in ee. Allele E^D was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. However, genotype frequencies of Limousin and Simmental breeds were 0.21 and 0.07 in E^DE^D, 0.00 and 0.03 in E^De^+ , 0.38 and 0.35 in E^De, respectively. These results of this study suggest that alleles of the bovine MC1R gene are associated strongly with coat color except for Simmental and Limousin breeds with dark brown, light yellow or mosaic coat color of red and brown. Therefore, it needs to be studied on relationship between A and E loci and coat color phenotypes. Furthermore, genotypes of the bovine MC1R gene can be used as a DNA marker for distinguishing. beef between Hanwoo and Holstein and Angus having black coat color phenotype.
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김태헌,윤두학,최승철,박응우,이영창,이장형,조진기 한국축산학회 1997 한국축산학회지 Vol.39 No.3
The purpose of this study was to investigate differences in restriction fragment length polymorphisms between mitochondrial DNAs(mtDNA) of Korean native pig and Landrace and to construct restriction map of the mtDNA of Korean native pig. When the mtDNA were digested with 16 different restriction enzymes of Apa I, Ava I, Barnes I, Bgl I, Bgl II, Cla I, EcoR I, EcoRV, Hind III, Hpa I, Pst I, Pvu II, Sac I, Sca I, Stu I and Xba I, to recognize 6 specific base pairs, restriction patterns of all the enzymes except for Bgl B and Sca I were identical between the two breeds. When digested with Bgl B and Sca I, seven out of ten Korean native pigs showed different band patterns from those of Landrace, but the other three showed the same patterns as those of Landrace. While five BamH I restriction fragments were reported in literature, the seven fragments were detected in this study. The restriction map of Korean native pig mitochondrial genome with 28 cleavage sites of 8 different restriction enzymes was constructed.
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