• Title/Summary/Keyword: Mitochondrial DNA

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Identification of Pork Adulteration in Processed Meat Products Using the Developed Mitochondrial DNA-Based Primers

  • Ha, Jimyeong;Kim, Sejeong;Lee, Jeeyeon;Lee, Soomin;Lee, Heeyoung;Choi, Yukyung;Oh, Hyemin;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.37 no.3
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    • pp.464-468
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    • 2017
  • The identification of pork in commercially processed meats is one of the most crucial issues in the food industry because of religious food ethics, medical purposes, and intentional adulteration to decrease production cost. This study therefore aimed to develop a method for the detection of pork adulteration in meat products using primers specific for pig mitochondrial DNA. Mitochondrial DNA sequences for pig, cattle, chicken, and sheep were obtained from GenBank and aligned. The 294-bp mitochondrial DNA D-loop region was selected as the pig target DNA sequence and appropriate primers were designed using the MUSCLE program. To evaluate primer sensitivity, pork-beef-chicken mixtures were prepared as follows: i) 0% pork-50% beef-50% chicken, ii) 1% pork-49.5% beef-49.5% chicken, iii) 2% pork-49% beef-49% chicken, iv) 5% pork-47.5% beef-47.5% chicken, v) 10% pork-45% beef-45% chicken, and vi) 100% pork-0% beef-0% chicken. In addition, a total of 35 commercially packaged products, including patties, nuggets, meatballs, and sausages containing processed chicken, beef, or a mixture of various meats, were purchased from commercial markets. The primers developed in our study were able to detect as little as 1% pork in the heat treated pork-beef-chicken mixtures. Of the 35 processed products, three samples were pork positive despite being labeled as beef or chicken only or as a beef-chicken mix. These results indicate that the developed primers could be used to detect pork adulteration in various processed meat products for application in safeguarding religious food ethics, detecting allergens, and preventing food adulteration.

Effect of Farnesyltransferase Inhibitor R115777 on Mitochondria of Plasmodium falciparum

  • Ha, Young Ran;Hwang, Bae-Geun;Hong, Yeonchul;Yang, Hye-Won;Lee, Sang Joon
    • Parasites, Hosts and Diseases
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    • v.53 no.4
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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.

Development of PCR Assay for Identification of Buffalo Meat

  • Rajapaksha, W.R.A.K.J.S.;Thilakaratne, I.D.S.I.P.;Chandrasiri, A.D.N.;Niroshan, T.D.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1046-1048
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    • 2003
  • A polymerase chain reaction (PCR) assay was developed to differentiate buffalo meat from the meat of Ceylon spotted deer (Axis axis ceylonensis), Ceylon sambhur (Cervus unicolor unicolor), cattle (Bovine), goat (Caprine), pig (Porcine), and sheep (Ovine). A set of primers were designed according to the sequence of the mitochondrial cytochrome b gene of bubalus bubalis and by PCR amplification a band of approximately 242 bp band was observed with buffalo DNA. These primers did not cross-react with DNA of other animal species tested in the study under the specified reaction conditions. A band of 649 bp was observed for all animal species tested when DNA was amplified with the universal primers indicating the presence of mitochondrial DNA in the samples. The technique was sensitive enough to identify rotten (10 days post slaughter), dried and cooked buffalo meat. The absence of a cross reaction with human DNA using the buffalo specific primers eliminates possible false positive reactions.

Analysis of Mitochondrial DNA Mutation in hepatoma

  • Chung, Ku-Sun;Lee, Kyo-Young;Shim, Sang-In;Kim, Jin-Sun;Song, Eun-Sook
    • BMB Reports
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    • v.33 no.5
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    • pp.417-421
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    • 2000
  • Mitochondrial DNA (mtDNA) mutation was investigated in a hepatoma patient using a polymerase chain reaction (PCR) and an in situ hybridization technique. Biotin-labeled probes for the subunit m of cytochrome c oxidase revealed differences in the in situ hybridization. A PCR assay using biopsied and microdissected tissues showed that common deletion (4,977 bp) was more pronounced in the cancer region than in the normal parts of the same patient. These results suggest that mtDNA deletion might be associated with tumorigenesis in hepatoma.

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Mitochondrial DNA analysis on 4 species of the genus Parus (Passeriformes: Paridae) in Korea (한국산 박새속 4종의 미토콘드리아 DNA 분석)

  • 민미숙
    • Animal Systematics, Evolution and Diversity
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    • v.13 no.2
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    • pp.73-81
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    • 1997
  • 한국산 조류 중 박새속(genus Parus)에 속하는 4종 Parus major wladiwostokensis (박새), P. ater amurensis(진박새), P. palustrius hellmayri(쇠박새) 및 P. varius varius(곤줄 박이)를 대상으로 이들의 계통적 유연관계를 구명하기 위하여 mtDNA분석을 실시하였다. 6 base를 인지하는 10개의 제한효소를 처리하여 얻어진 mtDNA의 크기는 16.6~17.0Kb였으며 Pst I과 Pvu II는 종간 차이가 뚜렷하였다. 각 종간의 절편양상을 비교하여 Parus속의 종 간 분화정도를 비교한 결과 P. m. wladiwostokensis와 P. a. amurensis의 유전적 근연관계 가 가장 가까웠고(p=0.073, F=0.294) P. a. amurensis와 P. v. varius는 비교적 현저한 유전 적 차이를 보였다.(p=0.119, F=0.143). Brown 등 (1979)의 공식을 이용하여 박새속 4종의 분 화시기를 추정한 결과 이들은 후기 선신세(Pliocene)와 초기 홍적세(Pleistocene) 사이에 분 화된 것으로 추정되었다.

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Sequence Comparison of Mitochondrial Small subunit Ribosomal DNA in Penicillium

  • Bae, Kyung-Sook;Hong, Soon-Gyu;Park, Yoon-Dong;Wonjin Jeong
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.62-65
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    • 2000
  • Partial sequence comparisons of mitochondrial small subunit rDNA (mt SSU rDNA) were used to examine taxonomic and evolutionary relationships among seven Penicillium species : two monoverticillate species, two biverticillate species, and three terverticillate species. Amplified fragments of mt SSU rDNA highly varied among seven species in size, suggesting the existence of multiple insertions or deletions in the region. A phylogengtic tree was constructed by exhaustive search of parsimony analysis. The phylogenetic tree distinguished two statistically supported monophyletic groups, one for two monoverticillate species and the other for three terverticillate species and ont biverticillate species, P. vulpinum. The phylogenetic relationship of P. waksmanii, the biverticillate species, was not clear.

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Nucleotide Sequence of 7.2 kb Mitochondrial Linear Plasmid DNA in Pleurotus ostreatus (Pleurotus ostreatus 미토콘드리아의 7.2 kb 선상 플라스미드 염기서열 분석)

  • 윤혜숙;구용범;노정혜
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.37-41
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    • 2001
  • Two linear plasmid-like DNAs, 10.2 kb and 7.2 kb were found in the mitochondria of P. ostreatus. They have covalently linked 5'-terminal proteins in both ends. Two continuous fragments of 4.7 kb and 2.3 kb from 7.2 kb DNA were cloned and sequenced. Two long open reading frames (ORF1; 2982 bp, 993 a.a and ORF2; 2703 bp, 900 a.a) and one short open reading frame(ORF3; 771 bp, 256 a.a) were found in the 7.2 kb plasmid. The putative ORF1 and ORF2 have conserved motifs of DNA polymerases and RNA polymerases, respectively, while the ORF3 has homologous regions with phosphatase from Plasmodium, and also with adhesine from Mycoplasma.

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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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.

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Association of ND4L gene 10609 mutation and hearing loss in a Korean with ESRD patients

  • Kim, Eun Sook
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.128-135
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    • 2012
  • The kidney and cochlea have similar physiological characteristics, specifically the active transport of fluid and electrolytes, similar effects of aminoglycosides and some immunological factors. Several mitochondrial DNA (mtDNA) defects have been identified to be associated with hearing impairment either in syndromic or nonsyndromic forms. Dialysis patients had more oxidative stress than healthy subjects and this elevated oxidative stress leads to alterations of the mtDNA. To generate a more comprehensive analysis of the relationship between mitochondrial variation and hearing loss, two SNPs of 10609, 14668 position showed nominal levels of association with hearing loss. In our result, the mean PTA values in the ESRD patients were $28{\pm}13.9\;(mean{\pm}SD)dB$ and $51.0{\pm}23.2dB$ in low and high frequencies, which were significantly higher than those in the normal controls. 10609T>C and 14668C>T were significantly associated with hearing loss in the ESRD patients. In summary, our results suggest that the polymorphisms of the ND4L subunit gene might be association with ESRD patients and hearing loss.

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Mitochondrial DNA Diversity of Korean Ogol Chicken

  • Lee, Y.J.;Bhuiyan, M.S.A.;Chung, H.J.;Jung, W.Y.;Choi, K.D.;Jang, B.G.;Paek, W.K.;Jeon, J.T.;Park, C.S.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.477-481
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    • 2007
  • Korean Ogol chicken has been registered as a natural monument in Korea and regarded as a valuable genetic resource for the world. As an initial step to investigate the genetic structures of this breed, phylogenetic analysis and calculation of genetic diversities have been performed using mitochondrial DNA (mtDNA) sequence variations. A total of 31 Korean Ogol chicken was grouped into four haplotypes and the large haplotype was represented in 12 individuals. The unrooted neighbor-joining tree indicates that the Korean Ogol chicken shared three (A to C) major chicken lineages representing the high genetic variability of this breed. These results can be used for making the breeding and conservation strategies for the Korean Ogol chicken.