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재래돼지 미토콘드리아 게놈내 D-loop 영역의 염기서열 분석
김태헌,윤두학,이효신,정일정,조진기 ( T . H . Kim,D . H . Yoon,H . S . Lee,I . C . Cheong,J . K . Jo ) 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.
김태헌,윤두학,최승철,박응우,이영창,이장형,조진기 ( T . H . Kim,D . H . Yoon,S . C . Choi,E . W . Park,Y . C . Lee,J . H . Lee,J . K . Jo ) 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.
한국 재래돼지 브랜드 돈육 원산지 검증을 위한 유전자 감식 기법 활용 연구
최봉암 ( Choi Bong-am ),이학교 ( Lee Hak-kyo ),전광주 ( Jeon Gwang-joo ),오재돈 ( Oh Jean-don ),최일신 ( Choi Il-sin ),박미현 ( Park Mi-hyun ),공홍식 ( Kong Hong-sik ),정일정 ( Jung Il-jung ),김태헌 ( Kim Tae-hun ),윤두학 ( Yoon D 한국유기농업학회 2004 韓國有機農業學會誌 Vol.12 No.2
Identification of animals has been used with an ear tag with dummy code and blood typing has been used for paternity and individual identification in live animals. Various genetic markers are different for breeds of pig and hence, it is necessary to identity the discrete genetic marker in korean native pig. A total of 240 pigs were used to find korean native pig population specific markers that expressed in population of korean native pigs. To identify the individual traceability, 20 animals were randomly chosen and tested for a whole process from being live to slaughter stages. The candidate genetic marker used in the study were 18 DNA microsatellites which were identified in pig genome. The number of alleles of those DNA microsatellites ranged form a minimum of 3 to maximum of 6. The heterozygote frequency ranged from 0.44 to 0.69. Effective number of alleles for each DNA microsatellotes were 2 to 4. By choosing 6 candidate genetic markers among all, the traceability of individual identification was estimated as accurate as 99.99%(p>0.0014), nearly.
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.
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.
장요순,윤두학,김태헌,정일정,조진기 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.2
ADRP 유전자가 24개월령 한우 등심조직에서 발현량이 급격히 증가하여 30개월령 등심조직에서는 발현량이 다소 감소하는 발현양상 분석결과로부터 이전 연구에서는 ADRP 유전자를 한우 성장단계 특이발현 유전자로 선정하였다. 본 연구에서는 ADRP 유전자의 발현조절 기작을 분석하기 위하여 promoter 영역을 포함하는 ADRP 유전자 전체영역을 cloning 하였으며, 구조를 분석하고 promoter의 특성을 조사하였다. 한우 ADRP cDNA 단편을 probe로 합성하여 Southern blot 분석을 실시한 결과로부터 ADRP 유전자가 한우 genome 상에서 single copy로 존재하고 크기는 대략 12kb에 해당하는 것을 확인하였다. Genomic DNA library screening을 실시하여 promoter 영역을 포함하는 ADRP 전체 유전자에 해당하는 clone을 확보하고 HwADRPg-1으로 명명한 후, 염기서열을 결정하고 분석하였다. 한우 ADRP 유전자, HwADRPg-1은 8개의 exon과 7개의 intron으로 구성되어 있으며 모든 exon-intron 경계는 GT/AG 원칙을 따르고 있었고, coding 영역은 7,633 bp로서 6개의 intron에 의해 7개의 exon으로 나누어져 있었다. HwADRPg-1의 promoter영역에서는 TATAA box는 발견되지 않았으며, -70 위치에 근육 특이적 transcription activator인 Myo G 서열이 존재하였고, -629 위치에는 지방세포의 분화를 유도하는 것으로 알려진 C/EBP (CCAAT/enhancer binding protein) 서열이 존재하였다. HwADRPg-1의 조절영역에 있는 Myo G factor가 근육조직에서 ADRP 유전자가 발현될 수 있도록 하며, 근육의 발달정도를 신호로써 감지하여 근육조직에서 성장단계에 따른 ADRP 유전자의 발현량을 조절할 것으로 추정되고, 다른 종류의 지방세포 특이적인 전사인자 및 지방세포의 분화정도를 신호로 인식하는 전사단계 조절인자를 조사하기 위하여 promoter 영역의 추가분석이 이루어져야 할 것으로 사료된다. To understand the structure and regulation of bovine ADRP (Adipocyte Differentiation Related Protein) gene, we have isolated the genomic clone of bovine ADRP and determined its sequence. A genomic Southern blot analysis confirmed that ADRP gene is present as a single copy in bovine genome and the ADRP gene spans 12 kb. Bovine ADRP genomic clone, HwADRPg-1, had 8 exons and 7 introns, and all splicing sites conformed to the GT/AG rule with the exon-intron boundaries located exactly. Analysis of the upstream 649 bp of the sequence of HwADRPg-1 showed that it does not contain any canonical TATAA boxes; however Sp1 binding sites and CAAT boxes are found. The promoter contained potential binding sites for AP-1, AP-2 and several putative transcription factor binding sites. The 5'-flanking region of HwADRPg-1 contained muscle specific transcription activator Myo G and C/EBP (CCAAT/enhancer binding protein) recognizing site. These results suppose that the Myo G transcription activator regulate the transcription of bovine ADRP gene in muscular tissue and its transcriptional activity was triggered by degree of muscular development. Our results provide the necessary analysis for other flanking sequences are needed in addition to the proximal cis elements of his promoter to confer adipocyte differentiation-dependent or growth-dependent transcriptional control.