• 환경생물
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• 제29권3호
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• pp.138-143
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• 2011

이번 연구에서는 중형저서동물의 생태학적 연구에 사용하는 Ludox와 Rose Bengal을 처리하였을 때, 고정액의 종류에 따른 DNA (mtCOI)의 추출 효율을 비교하고자 하였다. 실험을 위해 저서성 요각류 Tigriopus japonicus s.l.를 실험동물로 사용하였으며, 99% 에탄올과 4%포르말린의 두 종류의 고정액을 사용한 뒤 이들을 각각 대조구로 삼았다. 그리고 (1) Ludox HS40, (2) Rose Bengal, (3) Ludox HS40+Rose Bengal을 각각 시료와 반응시킨뒤, mtCOI 유전자를 추출하였다. 이후 PCR을 진행하고 산물을 전기영동하여 유전자의 증폭여부를 확인하였다. 또한 모든 실험은 30회 반복하여 실험간의 결과를 비교 하였다. 그 결과, 에탄올의 경우에는 대조구를 포함한 (1), (2), (3)의 실험 모두에서 96% 이상의 효율을 보였지만, 포르말린의 경우에는 대조구에서 27% 증폭되었으며, (1)과 (3)에서 약 3%, (2)에서 약 7%만 증폭되어 두고 정액에 따른 차이가 뚜렷하게 나타났다. 결과적으로 현재의 연구를 통해서 99% 에탄올이 중형저서동물에서 DNA를 추출하는 데 적합한 고정액임을 확인하였다.

• Journal of Preventive Medicine and Public Health
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• 제45권4호
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• pp.235-243
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• 2012

Objectives: Nail has been a substitute DNA source for genotyping. To investigate the integrity and consistency of nail DNA amplification for biomarker study, nail clippings from 12 subjects were collected at monthly intervals. The possibility of longer amplification and existence of GAPDH RNA/protein, were also investigated with three nail samples. Methods: Three primer sets were designed for quantitative amplification of nuclear and mitochondrial genes and analysis of their consistency. The mean threshold cycles in amplification of the target genes were compared to test the consistency of polymerase chain reaction (PCR) performance among individual factors including age groups, sex, family, the nail source, and by the size of the amplification segments. Results: The amplification of the target genes from nail DNA showed similar integrity and consistency between the nail sources, and among the serial collections. However, nail DNA from those in their forties showed earlier threshold cycles in amplification than those in their teens or seventies. Mitochondrial DNA (mtDNA) showed better DNA integrity and consistency in amplification of all three targets than did nuclear DNA (nucDNA). Over 9 kb of mtDNA was successfully amplified, and nested quantitative PCR showed reliable copy numbers (%) between the two loci. Reverse transcription PCR for mRNA and immunoblotting for GAPDH protein successfully reflected their corresponding amounts. Regarding the existence of RNA and protein in nails, more effective extraction and detection methods need to be set up to validate the feasibility in biomarker study. Conclusions: Nail DNA might be a feasible intra-individual monitoring biomarker. Considering integrity and consistency in target amplification, mtDNA would be a better target for biomarker research than nucDNA.

• 한국가금학회지
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• 제38권1호
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• pp.5-12
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• 2011

우리나라 재래닭의 기원을 구명하기 위해서 mtDNA D-loop 영역을 분석한 결과, 1,231~1,232개의 염기구성되어 있으며, 35개소에서 변이가 관찰되었다. 변이 부위를 이용하여 Haplotype을 분류한 결과, 21종으로 분류되었으며 이중 9개인 GenBank에 미등록된 것으로 밝혀졌다. Hplotype 다양성으로 추정한 한국 재래닭의 유전적 변이성은 중국의 재래닭과 유사한 것으로 추정되었다. Haplotype에 대한 Network 분석 결과, 재래닭은 5개의 Clade로 분류되었다. 이들 Clade에 대한 각 집단의 분포 현황으로 한국 재래닭은 운남성 및 중국 재래닭이 보인 결과와 유사하나, 일본의 재래닭과는 약간 상이한 것으로 밝혀졌다. 이상의 결과로 우리나라 재래닭은 공통선조가 다른 5개 이상의 모계가 중국을 통하여 유래되었으며, 일본에도 전파된 것이 확인되었다. 한편, 일본은 한반도를 유래하지 않은 닭의 유입이 있었던 것으로 추정된다.

• 한국환경과학회지
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• 제20권4호
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• pp.551-554
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• 2011

The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

• 생명과학회지
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• 제21권3호
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• pp.341-348
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• 2011

제주재래돼지의 모계 혈통에 대한 보다 명확한 이해를 얻기 위해, 본 연구에서는 제주재래돼지의 미토콘드리아 DNA (mtDNA) CYTB 유전자를 분석하고 이를 타 품종들에서 얻은 결과들과 비교하였다. 제주재래돼지를 포함한 돼지 6 품종에서 PCR-RFLP 분석을 수행하였고, RFLP 양상은 돼지 품종들을 뚜렷하게 구분되는 두 가지 반수체형(mtCYTB1 and mtCYTB2)으로 분리시켰다. 제주재래돼지 CYTB 서열들은 계통수 상에서 유럽과 아시아품종 cluster에서 모두 발견되었다. 제주재래돼지 CYTB들 중에서 J2 group은 중국재래돼지품종들과 근연이면서 아시아 고유 돼지 계통들과 함께 출현하였으며, 다른 한 group인 J1에 해당하는 서열들은 유럽돼지 계통들과 함께 위치하였고, 아시아 품종들보다는 스페인의 Iberian 재래돼지들과 근연인 것으로 확인되었다. 이 결과들은 현재 제주도에서 사육되고 있는 제주재래돼지 품종의 모계 기원은 크게 아시아계 돼지와 유럽계 돼지인 것으로 추정됨을 보여준다. 따라서 본 연구결과들은 제주재래돼지 집단은 과거에 가축화된 아시아 고유 돼지품종들과 공통 선조를 공유하고, 또한 20세기에 유입된 유럽계 돼지 품종들도 현재의 집단 형성에 기여한 것임을 시사하고 있다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권9호
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• pp.1223-1228
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• 2012

The population sizes of three Korean indigenous cattle populations have been drastically reduced over the past decades. In this study, we examined the extent to which reduction in populations influenced genetic diversity, population structure and demographic history using complete mitochondrial DNA (mtDNA) control region sequences. The complete mtDNA control region was sequenced in 56 individuals from Korean Black (KB), Jeju Black (JEB) and Korean Brindle (BRI) cattle populations. We included 27 mtDNA sequences of Korean Brown (BRO) from the GenBank database. Haplotype diversity estimate for the total population was high (0.870) while nucleotide diversity was low (0.004). The KB showed considerably low nucleotide (${\pi}$ = 0.001) and haplotype (h = 0.368) diversities. Analysis of molecular variance revealed a low level of genetic differentiation but this was highly significant (p<0.001) among the cattle populations. Of the total genetic diversity, 7.6% was attributable to among cattle populations diversity and the rest (92.4%) to differences within populations. The mismatch distribution analysis and neutrality tests revealed that KB population was in genetic equilibrium or decline. Indeed, unless an appropriate breeding management practice is developed, inbreeding and genetic drift will further impoverish genetic diversity of these cattle populations. Rational breed development and conservation strategy is needed to safeguard these cattle population.

• 보존과학연구
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• 통권25호
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• pp.47-74
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• 2004

We investigated the nucleotide substitution and insertion polymorphism of the hypervariable region Ⅰ and Ⅱ in mt DNA by sequencing ancient DNA from 51 ancient bones and teeth excavated at Nuk-do and Yimdang-dong in Korea. It revealed 35 sequence types from the ancient Korean. Of these, different sequences were 34 sequences. There were 19 and 38 base substitutions in HVI and HVⅡ, respectively. Some substitutions were characteristic of East Asian populations as compared with data reported on Caucacianpopulations,16051, 16150, 16172, 16223 in region I and 73, 263 in region II were noted as polymorphic sites, respectively. These were distributed evenly along the control region, though the frequency of each site was variable. Nucleotide substitution rather than insertion and deletion was the prevalent pattern of variation. Insertion of cytosine between312 and 315 in region HVⅡ were detected up to 98% in 51 ancient bone samples. This sequence data represents a phylogenetic tree using NTI DNA Suite computer program. The phylogenetic tree showed that mt DNA sequences of Nuk-do bones were relative to west Siberian and Indonesian. The usefulness of mt DNA sequencing in ancient Korean population excavated atarchaeological sites is based on biological and historical evidence for origin and migration of ancient Korean.

• 한국동물학회지
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• 제35권2호
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• pp.219-225
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• 1992

The genetic variation in mitochondrial DNA (mtDNA) was analysed within and between two species of tree frogs. Hyla japonica and H. suweonensis from South Korea. Purified mtDNAs were digested with each of 11 restriction enLvmes which cleave at six base recognition sequences. The genome size of H. iaponica revealed ho types (20.0 $\pm$ 0.3 and 19.6 $\pm$ 0.3 kb) and this difference is explained by either addition or deletion of about 0.4 kb fragment. On the other hand, the genome sire of H. suueonensis was about 19.0 $\pm$ 0.4 kb only. For the analysis, level of fragment homology (F) and nucleotide sequence divergence (p) were estimated from comparisons of digestion profiles. Among four populations of H. iaponica, substantial mean sequence divergence was 0.017 (range 0.001-0.026); between identical types, 0.001 IslilaRl type) and 0.004 (Large type) respectively; between different ones, 0.024 (range 0.023-0.026). The level of sequence divergence between he species was 0.142 (range 0.131-0.146). This result suggested that he species ㅂwere distinctly differentiated species. The divergence time between ko species was estimated 7.1 million years.

• Parasites, Hosts and Diseases
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• 제53권4호
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• pp.421-430
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• 2015

The parasite Plasmodium falciparum causes severe malaria and is the most dangerous to humans. However, it exhibits resistance to their drugs. Farnesyltransferase has been identified in pathogenic protozoa of the genera Plasmodium and the target of farnesyltransferase includes Ras family. Therefore, the inhibition of farnesyltransferase has been suggested as a new strategy for the treatment of malaria. However, the exact functional mechanism of this agent is still unknown. In addition, the effect of farnesyltransferase inhibitor (FTIs) on mitochondrial level of malaria parasites is not fully understood. In this study, therefore, the effect of a FTI R115777 on the function of mitochondria of P. falciparum was investigated experimentally. As a result, FTI R115777 was found to suppress the infection rate of malaria parasites under in vitro condition. It also reduces the copy number of mtDNA-encoded cytochrome c oxidase III. In addition, the mitochondrial membrane potential (${\Delta}{\Psi}m$) and the green fluorescence intensity of MitoTracker were decreased by FTI R115777. Chloroquine and atovaquone were measured by the mtDNA copy number as mitochondrial non-specific or specific inhibitor, respectively. Chloroquine did not affect the copy number of mtDNA-encoded cytochrome c oxidase III, while atovaquone induced to change the mtDNA copy number. These results suggest that FTI R115777 has strong influence on the mitochondrial function of P. falciparum. It may have therapeutic potential for malaria by targeting the mitochondria of parasites.

• 한국수정란이식학회지
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• 제18권3호
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• pp.227-235
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• 2003

한우의 mt DNA cytochrome oxidase subunit I, II, 및 III complex지역의 유전적 다형현상을 제한효소를 이용하여 검출하였다. PCR primer 6종에 대하여 20가지 제한효소를 처리하였으며, Pst I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I 제한효소를 사용하여 유전적 변이를 검출하였다. 검출된 변이체와 한우의 성장과의 관련성을 조사한 결과 cytochrome oxidase subunit III complex 지역의 유전염기서열을 근거로 제작한 primer Mt9 좌위에서 제한효소 PvuII를 이용한 절단형과 체중형질 인 WT15(P<0.05) 및 WT18(P<0.01)에서 고도의 유의성이 관찰되었다. 아울러 , Mt9-Pvu II(P=0.07), Mt6-Bgl II(P=0.05), and Mt8-Rsa I(P=0.05) 좌위 또한 WT9, WTl5, and WT15에서 각각 통계적 유의성이 관찰되었다. 따라서 본 결과는 cytochrome oxidase subunit III complex segments가 candidate gene으로서 기초적 유전정 보 제공은 물론 유전적 개량을 위해 사용될 수 있을 것으로 사료된다.