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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 2002 한국축산학회지 Vol.44 No.6
한우 성장단계 특이발현 유전자 탐색에 관한 연구(장 등, 2002)에서 선정된 후보유전자 중, EPV 20, TCTP 및 aldolase A를 비롯하여 24개월령 cDNA probe에 대하여 강한 signal을 나타낸 ADRP 유전자의 발현양상을 분석하기 위하여 성장단계별 한우 등심조직 시료를 사용하여 semiquantitative RT-PCR 및 northern blot 분석을 실시하였다. EPV 20 유전자는 한우 등심조직에서 성장단계에 따른 발현량 차이가 거의 없는 결과를 근거로, 근육조직에서 항상 발현되는 유전자로 판단하였다. 또한 발달단계에 따라 발현량 차이가 있는 것으로 보고된 aldolase A 유전자 역시 뚜렷한 발현량 차이는 없었으며, aldolase A의 muscle specific promoter와 근육분화 관련 transcription factor에 관한 연구를 통하여 근육분화 및 근육수축에 있어 aldolase A의 역할 및 조절작용을 밝힐 수 있을 것으로 사료된다. 성장관련 단백질로 알려져 있는 TCTP의 발현양상을 분석한 결과, 한우 등심조직에서 성장함에 따라 발현량이 약간 증가하는 양상을 나타내었다. 이와 같은 발현양상 분석결과로 TCTP 유전자는 성장과 관련된 기능이 있지만 동물의 성장에 있어 직접적으로 관여하지는 않을 것으로 판단된다. 지방세포 분화의 초기단계에서 발현량이 급증하고 lipid droplet 형성에 관여하며 지방축적과도 관련이 있는 것으로 보고된 ADRP 유전자의 transcript의 크기는 약 1.82 kb 이었고, 24개월령 한우 등심조직에서 발현량이 급격히 증가하였으며, 30개월령 조직에서는 감소하는 양상을 나타내었다. 이와 같은 결과로부터 ADRP 유전자를 한우 성장단계 특이발현 유전자로 성장하였으며, ADRP 유전자의 발현양상은 근육내 지방축적과 관련이 있을 것으로 추정하였다. 이후에는 발현조절 기작의 해명 및 근육내 지방축적과의 관련성 분석을 위하여 ADRP 유전자의 전체적인 구조 및 전사조절 인자 분석을 비롯하여 다른 지방대사 및 지방축적 관련 유전자와의 상호작용 및 다형성 탐색분석에 관한 연구가 필요 할 것으로 판단하였다. We have investigated the expression patterns of candidates for growth stage specifically expressed genes. The expression patterns of the EPV20, aldolase A, Translationally Controlled Tumor Protein (TCTP) and Adipocyte Differentiation Related Protein (ADRP) were examined by semiquantitative RT-PCR and northern blot analysis in skeletal muscle tissues of Hanwoo, especially in the longissimus dorsi at various growth stages. The EPV20 mRNA was expressed in longissimus dorsi tissue of Hanwoo, but there was no difference of expression levels during growth stages. Though the aldolase A gene was reported to be muscle-specific and regulated at developmental stages, the expression levels of aldolase A mRNA in the longissimus dorsi tissues showed little differences at various growth stages. The expression levels of TCTP which was reported as growth-related protein regulated at translation step were gradually increased during growth of Hanwoo. The expression levels of ADRP mRNA were rapidly increased at 24-month-old longissimus dorsi tissue of Hanwoo, and decreased at 30-month-old. Our data suggest that the ADRP gene show as growth-stage dependent expression and is related to fat deposition within muscular tissue.