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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.
약(葯)배양 기법 이용한 이탈리안 라이그라스 식물체 생산
김기용,강경민,최기준,임용우,장요순,성병렬,손대영,이병현,조진기 Plant molecular biology and biotechnology research 2002 Plant molecular biology and biotechnology research Vol.2002 No.-
We obtained regenerated Italian ryegrass (Lolium multiflorum Lam.) plants by anther culture. When Italian ryegrass anther was incubated for 20 days on callus induction medium, MS medium containing 30 g/ℓ of NAA and 1 mg/ℓ of kinetin its callus was induced. The ratio of callus induction was 9.2 %, the mean of callus weight was 8.6 mg/callus/anther. When Italian ryegrass callus was incubated for 50 days on plant regeneration medium, MS medium containing 30 g/ℓ of sucrose, 1 mg/ℓ of NAA and 2 mg/ℓ of kinetin, Italian ryegrass plant was regenerated. The ratio of plant regeneration was 26%.
Subtraction기법을 이용한 한우 성장 단계 특이 발현 유전자 탐색
장요순,김태헌,윤두학,정일정,조진기,박응우 한국동물자원과학회 2002 한국축산학회지 Vol.44 No.1
한우의 성장단계 특이발현 유전자를 탐색하기 위하여, 본 연구에서는 유전자의 발현 유무 및 발현정도의 차이를 나타내는 유전자를 분리하는데 있어 가장 강력한 수단으로 알려진 subtractive cDNA hybridization 기법을 이용하여 한우 등심조직으로부터 12개월령 및 24개월령 특이적인 subtractive cDNA library를 제작하였다. 성장단계 특이적인 유전자를 탐색하기 위하여, 6, 12 및 24개월령 cDNA를 사용하여 reverse northern blot 분석을 실시하였으며, 6개월령 cDNA probe에 대하여 특이적인 signal을 나타낸 3개의 clone은 EPV 20, Ca^2+ ATPase, 및 TCTP 유전자와 유사성을 나타내었다. 12개월령 cDNA probe에 대하여 특이적인 signal을 나타낸 9개의 cDNA clone은 각각 VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase T1 및 UCP2 유전자와 높은 homology를 가지고 있었다. 또한 2개의 clone이 각각 12개월령 및 24개월령 cDNA probe에 대하여 특이적인 signal을 나타내었는데, 12개월령 cDNA probe에 대해서만 signal을 나타낸 clone은 ferrochelatase 유전자와 유사하였으며, 24개월령 probe에 대해서만 signal을 나타낸 clone은 ADRP 유전자와 유사하였다. 이상에서와 같이, 본 연구에서 제작한 성장단계 특이적인 subtractive cDNA library를 분석하여 14종의 유전자를 한우 성장단계 특이 발현 후보 유전자로 선정하여 염기서열을 분석하였으며, 이외에도 성장단계에 있어 특이적으로 발현될 것으로 추정되는 cDNA 클론의 염기서열을 분석하였다. To identify the differentially expressed genes at growth stage of Hanwoo, we constructed the subtractive cDNA library from loin mRNA of 12- and 24-month old Hanwoo by PCR-based subtraction. The fourteen genes were confirmed by sequencing and reverse northern blot analysis, and they were selected as candidate of putative genes differentially expressed at the growth stage of Hanwoo. Three subtracted cDNA fragments that expressed specific signal to cDNA probe for 6-month-old loin of Hanwoo were highly homologous to those of the genes encoding EPV 20, Ca^2+ ATPase, and TCTP, respectively. The nine cDNA clones showed intense signal to cDNA probe from 12-month-old loin of Hanwoo, and highly homologus to those of genes encoding VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein pl, ADP/ATP translocase, and UCP2, respectively. Two subtracted cDNA clones that expressed specific signal to cDNA probes for 12- and 24-month-old loin of Hanwoo were detected. One of them was highly homologus to the gene encoding ferrochelatase and the other was highly homologus to the gene encoding ADRP.
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.