• Animal Systematics, Evolution and Diversity
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• 제11권4호
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• pp.469-477
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• 1995

Drosophila virilis section 중 D. robusta 종군내에서 D. lacertosa 아군의 D. lacertosa 와 D. robusta 아군의 D.sordidula 에 대한 형태학적 특징 및 세포학적 특징(핵형분석)간에 차이가 크기 때문에 이 두종을 대상으로 종의 분화정도와 종형성 과정을 알아보기 위한 연구의 일환으로 10가지 제한효소를 사용하여 mtDNA 의 절단인식부위를 분석하였다. 그 결과, D.lacertosa 와 D.sordidula 의 전체 genome size는 공히 15.7kbp 인 것으로 나타났으며, mtDNa 의 제한효소 fragment 수는 각각 26개와 32개인 것으로 조사되었다. 또한 2 종류의 제한효소를 동시에 처리하여 제한효소 지도를 작성하여 보았을 때, 이들 두종의 제한효소 지도의 형태에서 매우 큰 차이가 있는 것으로 나타났다. 같은 종군내에서 형태학적 , 세포학적 차이 및 mtDNA 의 제한효소 지도에 의한 차이로 볼 때 이들 2종의 차이는 아군수준으로까지 분류할 수 있을정도로 두종의 분화가 오래전에 이루어졌음을 시사하였다.

• Animal cells and systems
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• 제3권1호
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• pp.73-78
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• 1999

Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

• Journal of Ginseng Research
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• 제14권2호
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• pp.310-316
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• 1990

This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular 9enetical approach of energy Production related mechanism in Panax ginseng. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. mtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not. Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRII treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN I and EcoRII. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector. Genomic library was screened and purified for further research including restriction mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned from the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

• 대한수의학회지
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• 제34권2호
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• pp.243-247
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• 1994

As a result of the detection of mitochondrial DNA(mtDNA) polymorphism to Thoroughbred and Percheron using 14 restriction enzymes, mtDNA polymorphism of Cheju horse observed in the Bam HI and Sac I. Only in both restriction enzymes two types were classified as of A type, which is high expression frequency and B type, which is low expression frequency. In the other 12 restriction enzymes mtDNA polymorphism was not detected. On the basis of this information mtDNA polymorphism of Cheju horse was examined but was not observed the polymorphism and only A type was expressed both Bam HI and Sac I restriction enzymes. Through this study Cheju horse was demonstrated that lower genetic variation was expressed from the detection of mtDNA polymorphism.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
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• pp.168-174
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• 1990

This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular genetically approach of energy Production related mechanism in Panax Ein.fend. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. MtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRll treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN 1 and EcoRll. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector Genomic library was screened and purified for further research including restricttion mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned koto the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

• Parasites, Hosts and Diseases
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• 제33권4호
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• pp.331-340
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• 1995

형태적으로 Aconthamoeba polyphaga로 동정긴 일곱 분리주의 동위효소 profile과 Mitochondria (Mt) DNAangerprint를 비교 분석하였다. 8가지 제한효소(B91 II. Sca I, Cla I, EcoR I, Xba I, Kpa I, Sal I. 및 Sst I)에 의한 Mt DNA fhlgerprint는 주간의 심한 다양성을 나타내었다. B가지 동위효소 (acid phosphatase, lactate dehydrogenase 및 glucose-6-phosphate dehydrogenase)는 주간의 심한 다양성을 나타내었으나 다른 3가지 동위효소(glucose phosphate isomerase, leucine aminopeptidase 및 malatedehydrogenase)는 비슷한 양상으로 나타났다 Ap 주의 동위효소 양상과 Mt DNA fmgerprint는 Jones 주와 동일하였다. Mt DNA fingerprinting은 대식가시아메바 주의 동정과 분리에 매우 유용함을 알았다. Aconthamoeba polvphasa의 우리말 학명을 대식가시아메바로 제안한다.

• Journal of Radiation Protection and Research
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• 제34권2호
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• pp.49-54
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• 2009

Mitochondrial DNA (mtDNA) deletion is a well-known marker for oxidative stress and aging and also contributes to their unfavorable effects in cultured cells and animal tissues. This study was conducted to investigate the effect of ionizing radiation (IR) on mtDNA deletion and the involvement of reactive oxygen species (ROS) in this process in human lung fibroblast (IMR-90) cells. Young IMR-90 cells at population doubling (PD) 39 were irradiated with $^{137}Cs$ $\gamma$-rays and the intracellular ROS level was determined by 2',7'-dichlorofluorescein diacetate (DCFH-DA) and mtDNA common deletion (4977bp) was detected by nested PCR. Old cells at PD 55 and $H_2O_2$-treated young cells were compared as the positive control. IR increased the intracellular ROS level and mtDNA 4977 bp deletion in IMR-90 cells dose-dependently. The increases of ROS level and mtDNA deletion were also observed in old cells and $H_2O_2$-treated young cells. To confirm the increased ROS level is essential for mtDNA deletion in irradiated cells, the effects of N-acetylcysteine (NAC) on IRinduced ROS and mtDNA deletion were examined. 5 mM NAC significantly attenuated the IR-induced ROS increase and mtDNA deletion. These results suggest that IR induces the mtDNA deletion and this process is mediated by ROS in IMR-90 cells.

• Journal of Radiation Protection and Research
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• 제36권3호
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• pp.119-126
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• 2011

Mitochondrial DNA (mtDNA) deletion is a well-known marker for oxidative stress and aging, and contributes to harmful effects in cultured cells and animal tissues. mtDNA biogenesis genes (NRF-1, TFAM) are essential for the maintenance of mtDNA, as well as the transcription and replication of mitochondrial genomes. Considering that oxidative stress is known to affect mitochondrial biogenesis, we hypothesized that ionizing radiation (IR)-induced reactive oxygen species (ROS) causes mtDNA deletion by modulating the mitochondrial biogenesis, thereby leading to cellular senescence. Therefore, we examined the effects of IR on ROS levels, cellular senescence, mitochondrial biogenesis, and mtDNA deletion in IMR-90 human lung fibroblast cells. Young IMR-90 cells at population doubling (PD) 39 were irradiated at 4 or 8 Gy. Old cells at PD55, and H2O2-treated young cells at PD 39, were compared as a positive control. The IR increased the intracellular ROS level, senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) activity, and mtDNA common deletion (4977 bp), and it decreased the mRNA expression of NRF-1 and TFAM in IMR-90 cells. Similar results were also observed in old cells (PD 55) and $H_2O_2$-treated young cells. To confirm that a increase in ROS level is essential for mtDNA deletion and changes of mitochondrial biogenesis in irradiated cells, the effects of N-acetylcysteine (NAC) were examined. In irradiated and $H_2O_2$-treated cells, 5 mM NAC significantly attenuated the increases of ROS, mtDNA deletion, and SA-${\beta}$-gal activity, and recovered from decreased expressions of NRF-1 and TFAM mRNA. These results suggest that ROS is a key cause of IR-induced mtDNA deletion, and the suppression of the mitochondrial biogenesis gene may mediate this process.

• Animal Systematics, Evolution and Diversity
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• 제13권1호
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• pp.21-27
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• 1997

한국 고유종인 새코미꾸리(Cobitis rotundicaudata)의 집단간 유전적 차이에 따른 종 분화 여부를 밝히고자 4개집단을 대상으로mitochondrial DNA(mtDNA)의 RFLP분석을 실시 하였다. C. rotundicaudata mtDNA를 10개의 6-base cutting 제한요소로 처리한 다음 그 절 편 양상을 비교, 분석한 결과 4개 집단 공히 mtDNA의 전체 genome 크기는 약 16.5$\pm$ 0.5Kbp였으며 공통절편수(F)에서 한강 2개 집단(가평, 진부)과 동해안 마읍천 짐단간의 F값 은 0.911로 매우 가까웠으나 낙동강의 산청집단은 타 3개 집단과 F=0.375로 차이가 있었다. 또한 염기치환율(p)에 있어서도 한강 2개 집단 및 마읍천 집단간은 평균 p=0.005로 매우 유 사하였으나, 산청 집단은 타 집단들과 염기치환율에 있어 p=0.059로 종수준의 뚜렷한 차이 를 나타내어서 이들은 각각 별종으로 사료된다.

• 한국수산과학회지
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• 제30권5호
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• pp.804-808
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• 1997

한국산 참굴의 유전적 특성을 조사하기 위하여 한국의 지역별 참굴을 대상으로 mtDNA 제한효소 절편분석과 클로닝을 수행하였다. 지역별로 각각 20개체의 mtDNA에 대하여 8가지 제한효소를 사용하여 DNA 절편양상을 분석한 결과 서해안산 참굴에서는 개체간 차이가 없는 단일 양상이었으며 남해안산의 경우는 두 가지 양상을 보였으며 차이는 HindIII 절단양상에서만 나타났다. 그 중 소수 개체들에서 나타난 양상은 서해안산 개체들에서의 양상과 동일하여 남해안에 서해안산 참굴이 유입되어 혼재하는 것으로 추정되었다. 한국산 참굴의 mtDNA를 대장균 E. coli HB101에서 클로닝하여 유전적 분석을 용이하도록 하였다. 전체 mtDNA를 제한효소를 사용하여 세부분으로 나누어 pUC19 유전자운반체에 클로닝하였다. 클로닝된 재조합 DNA를 제한효소들로 절단하여 한국산 참굴 mtDNA의 제한효소지도를 작성하였다. 남해안산과 서해안산 참굴 mtDNA에서 HindIII 적단 양상이 다르게 나타남을 확인하였고 이는 남해안산이나 서해안산에서 염기치환의 돌연변이에 의한 것으로 사료되었다.