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해수에서 분리한 Vibrio sp. M-96 균주의 열감수성 alkaline phosphatase 성질
박문경,진덕희,김중균,공인수,김광현,홍용기,Park, Moon-Kyung,Jin, Deuk-Hee,Kim, Joong-Kyun,Kong, In-Soo,Kim, Kwang-Hyeon,Hong, Yong-Ki 한국생명과학회 1996 생명과학회지 Vol.6 No.3
A thermolabile alkaline phosphatase has been purified through steps of osmotic shock, ammonium sulfate salting-out, and DAEA-cellulose chromatography from the cultured broth of the marine Vibrio sp. M-96 strain. The optimal temperature for the enzyme activity was 35$\circ$C. The optimal pH was pH11.0, and the range of pHstability was pH10.4 to 12.0. Thermal inactivation occured within 6 mintes at 60$\circ$C. The enzyme was considerably inactivated by 0.1mM concentrations of Hg$^{2+}$, Ni$^{2+}$ and Zn$^{2+}$, whereas activated up to 234% by 1mM of Mn$^{2+}$. The activation energy and deactivation energy by the Arrhenius equation were 4.02 Kcal/mol and 9.098 Kcal/mol, respectively. The Km and Vmax values of the enzyme for p-introphenylphosphate were found to be 0.0465mM and 0.001334mM/min, respectively. Active form of the enzyme had a molecular weight of 57,000 dalton determined by the Sephadex G-200 gel filtration method.
넙치와 조기의 원산지 판별을 위한 random amplified polymorphic DNA 패턴 연구
강덕진,이석근,진덕희,최석정,Kang Duk-Jin,Lee Suk-Keun,Jin Deuk-Hee,Choi Suk-Jung 한국생명과학회 2006 생명과학회지 Vol.16 No.1
본 연구에서 넙치와 조기의 원산지를 판별하기 위한 도구로 RAPD PCR 방법의 가능성을 확인하였다. 넙치는 한국의 주문진 자연산, 통영 양식산, 거제 양식산 그리고 북한 자연산을 실험에 사용하였다. 조기는 한국산과 중국산을 사용하였다. 넙치의 RAPD 패턴에서는 뚜렷하고 일관성이 있는 진단용 띠들을 쉽게 찾을 수 있었다. 조기의 경우에는 유전적인 이질성으로 인하여 각 개체의 RAPD 패턴에서는 일관성이 있는 진단용 띠를 찾기 어려웠지만 각 원산지별로 얻은 RAPD 패턴에서는 가능성이 있는 진단용 띠들을 찾을 수 있었다. The random amplified polymorphic DNA (RAPD) technique was investigated as a potential tool for the origin identification of olive flounder (Paralichthys olivaceus) and redlip croaker (Pseudosciaena polyactis). Olive flounder specimens were collected from North Korea and several locations of South Korea (Jumunjin, Tongyoung and Geoje). Fishes obtained from Tongyeong and Geoje were cultured products. Redlip croaker specimens were collected from South Korea and China. Consistent and distinct diagnostic bands were easily identified in the RAPD patterns of the olive flounder specimens. Although consistent diagnostic bands rarely appeared in the RAPD pattern of redlip croaker specimens because of their genetic heterogeneity, we were able to find potential diagnostic bands in the average RAPD pattern of each origin.
Denaturing gradient gel electrophoresis 와 real time PCR 방법을 이용한 연어 유전자들의 DNA 이형 다양성 검색
함승협(Seung Hyup Ham),이석근(Suk Keun Lee),한현섭(Hyon Sob Ham),진덕희(Deuk Hee Jin) 한국수산과학회 2002 한국수산과학회지 Vol.35 No.5
한국, 미국, 일본지역에 서식하는 연어에서 추출한 genomic DNA를 이용하여 연어의 mtDNA NDI 영역, D-loop 영역, growth hormone, IGF-I, MCH2, histone H3의 염기서열을 분석하여, 최적의 primer를 제작하여 PCR을 실시한 결과, mtDNA ND1 영역은 Ks12, Ks24, As11, As14, Js13, Js15에서 증폭된 DNA를 확인하였으며, D-loop 영역, growth hormone, IGF-I, histone H3, MCH2에서는 모든 시료에서 증폭된 DNA를 확인하였다. DGGE 분석의 결과, mtDNA NDI 영역 (AF133701, 449-880), D-loop 영역 (AF125518, 11-514)과 growth hormone (AF005927, 181-530)에서는 이형다양성을 확인하였으며, IGF-I (AF063216, 996-1461)과 MCH2 (M2728l, 70-593)는 모두 이형다양성이 나타났으나, histone H3 (AF017147, 7-487)는 모두 이형다양성이 관찰되지 않았다. 그리고 real time PCR 관찰 결과는 DGGE의 결과와 유사한 점을 찾을 수 없었지만, real time PCR도 각각의 유전자에 따라 서로 다른 DNA 생성 패턴을 보여 DNA 변이를 쉽게 구별하는데 보조적인 도움이 되었다. In order to detect the DNA heteropolymorphism of chum salmon, selected essential genes were examined in different regional chum salmons, i.e., Korean, Japanese and American by denaturing gradient gel electrophoresis (DGGE) and real time PCR methods. From the promoter regions and introns of growth hormone, mtDNA ND1 region, D-loop region, IGF-I, histone H3 and MCH2 several representative primer pairs were obtained and employed for the DGGE with the PCR products from the genomic DNAs of the different regional chum salmons. mtDNA ND1, D-loop region and IGF-I genes showed marked heteropolymorphism between Korean and American chum salmons. Intron C of growth hormone also showed a heteropolymorphism between Korean and Japanese chum salmons. Whereas heteropolymorphism of histone H3 and MCH2 genes was detected among in Korean, Japanese and American chum salmons in the examined region. The real time PCR disclosed the characteristic incremental production of target DNAs dependent on the heteropolymorphic conditions of genomic DNAs of chum salmons, thus the different regional chum salmons could be grouped by the variable incremental curves. Although the DGGE and real time PCR did not produce the identical results in this study, we suggest that the DGGE and real time PCR could be used for the primary screening of the DNA heteropolymorphism of different animal genome.
한국산 연어의 미토콘드리아 NADH Dehydrogenase Subunit 3 영역의 클로닝 및 DNA 염기서열 분석
최윤실 ( Yoon Sil Choi ),이윤호 ( Youn Ho Lee ),진덕희 ( Deuk Hee Jin ) 한국수산과학회 2003 한국수산과학회지 Vol.36 No.2
N/A Mitochondrial DNAs has been used frequently as genetic markers for the population genetic studies of salmonid fishes. Samples used in this experiment were chum salmons (Oncorhynchus keta) from Korea. We analyzed variation of mitochondrial NADH dehydrogenase subunit 3 gene (ND3) among 4 individuals of the Korea population. Genomic DNA was extracted from the liver of the chum salmon samples. Then, the ND3 gene was amplified by polymerase chain reaction (PCR) including the 3`region of cytochrome oxidase III gene (COIII) and the 5`region of NADH dehydrogenase subunit 4L gene (ND4L). The size of the PCR product was 752 bp and the sequences showed some genetic variation among those four individuals. Genetic variations were observed in 7 sites as single nucleotide polymorphism (SNP). Within the open reading frame of ND3 gene which encodes 116 amino acids, 5 nucleotide substitutions were found. Both transitional and the others of a Japanese chum salmon in Genbank showed 5 sites of SNPs. This study provided the basic information of SNP in ND3 gene among Korean chum salmons and demonstrated the possible use of the SNP data as a genetic marker.
Jun Gyo Jung(정준교),Jae Hong Kim(김재홍),Jeong Hwan Kim(김정환),Yong Soo Kim(김용수),Deuk-Hee Jin(진덕희),Hyung-Joo Jin(진형주) 한국생명과학회 2021 생명과학회지 Vol.31 No.8
생활습관 개선을 통한 근육량향상은 대사 증후군의 위험을 줄이는 것으로 보고되었다. 본 연구에서는 갈조류인 고리매(Scytosiphon lomentaria) 에탄올 추출물이 골격근 성장억제조절 단백질인 마이오스타틴 신호전달을 억제하는지와 고 급식으로 유도된 비만 제브라피시의 대사 항상성에 대한 효과를 확인하였다. 고리매 에탄올 추출물(10 ㎍/ml)은 pGL3- (CAGA) 12-루시퍼라제 분석에서 마이오스타틴(1 nM/ml) 신호를 완전히 차단하였다. 또한 웨스턴 블롯 분석에서 마이오스타틴 신호를 차단하여 Smad2 인산화가 억제되는 것을 확인하였다. 제브라피쉬의 치어에 대한 연구는 고 급식 대조군 그룹의 체내 포도당 농도는 정상 급식 대조군 그룹보다 유의하게 높았지만, 12.5 ㎍의 고리매 에탄올을 처리한 고 급식 그룹과 18.75 ug의 고리매 에탄올로 처리한 고 급식 그룹의 체내 포도당 수준은 정상 급식 대조군 그룹과 유사하였다. 고 급식 그룹의 GLUT2 유전자의 mRNA 발현 수준은 정상 급식 대조군 그룹에 비해 현저히 낮았다. 하지만, 고리매 에탄올 추출물을 처리한 실험군 그룹의 GLUT2 유전자의 발현은 정상 급식 대조군 그룹의 GLUT2 유전자의 발현과 거의 유사하게 나타났다. 그러므로 고리매 에탄올 추출물은 GLUT2유전자의 발현 조절을 통하여 체내 포도당 조절이 가능함을 보여준다. 본 연구의 결과는 고리매 에탄올 추출물이 대사 증후군 치료에 도움을 주는 소재 및 마이오스타틴 억제제로서의 가능성을 시사합니다. Muscle mass improvement through lifestyle modification has been shown to reduce the risk of metabolic syndrome. This study examined the capacity of ethanol extracts of Scytosiphon lomentaria (SLE) to suppress the bioactivity of myostatin, a potent negative regulator of skeletal muscle mass, as well as the effect of SLE treatment on metabolic homeostasis in obese zebrafish induced by high feeding. A total of 10 ㎍/ml SLE completely blocked myostatin (1 nM/ml) signaling in the pGL3-(CAGA)12 luciferase assay and suppressed myostatin-induced Smad2 phosphorylation in the Western blot analysis. In the zebrafish larvae analysis, the whole body glucose concentration of the high feeding control (HFC) group was significantly higher than that of the normal feeding control (NFC) group. However, the glucose levels of the high feeding group treated with 12.5 ㎍ SLE and of the high feeding group treated with 18.75 ug SLE were similar to those of the NFC group. The mRNA expression level of the GLUT2 gene of the HFC group was significantly lower than that of the NFC group. SLE treatment restored the expression of the GLUT2 gene to a level that was close to that of the NFC group, indicating that SLE is capable of regulating glucose levels in zebrafish larvae. The current results highlight the potential of SLE as a natural MSTN inhibitor and supplement that can be used to facilitate the treatment of metabolic syndrome.