• Title, Summary, Keyword: mitochondrial DNA

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Mitochondrial DNA Analysis in Fusants of Ganoderma lucidum and Lentinus edodes (영지버섯과 표고버섯 원형질 융합체의 미토콘드리아 DNA 검색)

  • 최은주;정영자;이영재;김병각;현진원
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.199-204
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    • 2002
  • It has been known that Ganoderma lucidum and Lentinus edodes have anticancer activity and immune enhancing activity. These two mushrooms were grown in liquid culture and harvested. From these mycelia, DNA was isolated and EtBr-CsCl density gradient ultracentrifugation was performed to purify it further. Then mitochondrial DNA was isolated by bisbenzimide-CsCl density ultracentrifugaton. Mitochondrial DNA of Ganoderma lucidum was digested by restriction enzymes, EcoR I, Hind Ⅲ, and Pst I, then electrophoresed. It showed 12, 22, 4 fragments. Mitochondrial DNA of Lentinus edodes was digested by EcoR I. Electric pattern showed 6 fragments. 4 fragments had appeared by Pst 1 digested mitochondrial DNA. Hind ill couldn't digest mitochondrial DNA of Lentinus edodes. Mitochondrial DNA of fusants was isolated to compare to those of parents. The results showed that fusant P₂S₄has new, recombined mitochondrial DNA. But P₂S₄had the same DNA that Ganoderma lucidum had.

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MITOCHONDRIAL DNA DELETION AND IMPAIRMENT OF MITOCHONDRIAL BIOGENESIS ARE MEDIATED BY REACTIVE OXYGEN SPECIES IN IONIZING RADIATION-INDUCED PREMATURE SENESCENCE

  • Eom, Hyeon-Soo;Jung, U-Hee;Jo, Sung-Kee;Kim, Young-Sang
    • Journal of Radiation Protection and Research
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    • v.36 no.3
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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.

Restriction and transcription maps of mitochondrial DNA of trimorphomyces papilionaceus

  • Jeoung, Won-Jin;Hong, Soon-Gyu;Won, Kang-Young;Jung, Hack-Sung
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.149-153
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    • 1995
  • Mitochondrial DNA has been isolated from Trimorphomyces papilionaceus. By analyzing DNA fragments digested by restriction enzymes, a restriction site map has been constructured. The mtDNA of T. papilionaceus amounts to 48.5 kb in size and is circular in structure. Entire mitochondrial DNA was cloned in E coli plasmids and Northern blot hybridization was done using cloned and subcloned DNAs as probes. Based on hybridization results of mitochondrial RNA transcripts, a transcription map was prepared.

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Mitochondrial DNA Mutation and Oxidative Stress

  • Kim, Tae-Ho;Kim, Hans-H.;Joo, Hyun
    • Interdisciplinary Bio Central
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    • v.3 no.4
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    • pp.16.1-16.8
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    • 2011
  • Defects in mitochondrial DNA (mtDNA) cause many human diseases and are critical factors that contribute to aging. The mechanisms of maternally-inherited mtDNA mutations are well studied. However, the role of acquired mutations during the aging process is still poorly understood. The most plausible mechanism is that increased reactive oxygen species (ROS) may affect the opening of mitochondrial voltage dependent anion channel (VDAC) and thus results in damage to mtDNA. This review focuses on recent trends in mtDNA research and the mutations that appear to be associated with increased ROS.

Development of a Highly Efficient Isolation Protocol for Mitochondrial DNA and RNA Using Small Scale Plant Tissues (식물의 초경량 조직을 이용한 미토콘드리아의 DNA와 RNA 정제)

  • Kim Kyung-Min;Lim Yong-Suk;Shin Dong-Ill;Sul Ill-Whan
    • Journal of Life Science
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    • v.16 no.2
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    • pp.240-244
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    • 2006
  • We present a fast and simple protocol for purification of mitochondria, mitochondrial DNA, and RNA from small amounts of tomato leaves. This method uses a high ionic strength medium to isolate mitochondria and extract mitochondrial DNA and RNA from a single preparation and is easily adaptable to other plant species. Mitochondria was confirmed by MitoTracker. The mitochondrial DNA was not contaminated by plastid DNA, was successfully used for PCR. Similarly, the isolated mitochondrial RNA was not contaminated only slightly contaminated (leaves) by plastid RNA. RNA prepared according to our method was acceptable for RT-PCR analysis

Mitochondrial DNA Polymorphism, Maternal Lineage and Correlations with Postnatal Growth of Japanese Black Beef Cattle to Yearling Age

  • Malau-Aduli, A.E.O.;Nishimura-Abe, A.;Niibayas, T.;Yasuda, Y.;Kojima, T.;Abe, S.;Oshima, K;Hasegawa, K.;Komatsu, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1484-1490
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    • 2004
  • Mitochondrial DNA haplotypes from the displacement-loop (D-loop) region (436 bp) were genotyped and sequenced in Japanese Black beef cattle raised in the same herd. Correlation coefficients between mitochondrial DNA haplotypes, maternal lineage, birth weight, preweaning average daily gain, weaning weight, post weaning average daily gain and yearling weight were computed. The objective was to study the relationship between maternal and postnatal growth traits and to investigate if postnatal growth of calves to yearling age could be accurately predicted from mitochondrial DNA haplotypes. Results of the phylogenetic analysis revealed 17 maternal lineages and four mitochondrial DNA haplotypes. There were strong, positive and highly significant (p<0.001) correlations among maternal traits ranging from 0.52 to 0.98. Similarly, among postnatal growth traits, most of the correlations were also strong, positive and highly significant (p<0.001); the highest correlation of 0.94 was between preweaning average daily gain and weaning weight. However, correlations between mitochondrial DNA haplotypes and postnatal growth traits were very low, mostly negative and non-significant (p>0.05) ranging from -0.05 to 0.1. Prediction of postnatal growth from mitochondrial DNA yielded very low $R^{2}$ values ranging from 0.002 to 0.019. It was concluded that mitochondrial DNA polymorphism has no significant association with postnatal growth from birth to yearling age, and by implication, nuclear rather than cytoplasmic DNA, accounts for most of the genetic variation observed in postnatal growth of Japanese Black cattle. Therefore, mitochondrial DNA genotyping at an early age has no bearing on the accurate prediction of the future growth performance of calves.

Analysis of Nuclear Mitochondrial DNA Segments of Nine Plant Species: Size, Distribution, and Insertion Loci

  • Ko, Young-Joon;Kim, Sangsoo
    • Genomics & Informatics
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    • v.14 no.3
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    • pp.90-95
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    • 2016
  • Nuclear mitochondrial DNA segment (Numt) insertion describes a well-known phenomenon of mitochondrial DNA transfer into a eukaryotic nuclear genome. However, it has not been well understood, especially in plants. Numt insertion patterns vary from species to species in different kingdoms. In this study, the patterns were surveyed in nine plant species, and we found some tip-offs. First, when the mitochondrial genome size is relatively large, the portion of the longer Numt is also larger than the short one. Second, the whole genome duplication event increases the ratio of the shorter Numt portion in the size distribution. Third, Numt insertions are enriched in exon regions. This analysis may be helpful for understanding plant evolution.

Mechanisms of Uniparental Mitochondrial DNA Inheritance in Cryptococcus neoformans

  • Gyawali, Rachana;Lin, Xiaorong
    • Mycobiology
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    • v.39 no.4
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    • pp.235-242
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    • 2011
  • In contrast to the nuclear genome, the mitochondrial genome does not follow Mendelian laws of inheritance. The nuclear genome of meiotic progeny comes from the recombination of both parental genomes, whereas the meiotic progeny could inherit mitochondria from one, the other, or both parents. In fact, one fascinating phenomenon is that mitochondrial DNA in the majority of eukaryotes is inherited from only one particular parent. Typically, such unidirectional and uniparental inheritance of mitochondrial DNA can be explained by the size of the gametes involved in mating, with the larger gamete contributing towards mitochondrial DNA inheritance. However, in the human fungal pathogen Cryptococcus neoformans, bisexual mating involves the fusion of two isogamous cells of mating type (MAT) a and MAT${\alpha}$, yet the mitochondrial DNA is inherited predominantly from the MATa parent. Although the exact mechanism underlying such uniparental mitochondrial inheritance in this fungus is still unclear, various hypotheses have been proposed. Elucidating the mechanism of mitochondrial inheritance in this clinically important and genetically amenable eukaryotic microbe will yield insights into general mechanisms that are likely conserved in higher eukaryotes. In this review, we highlight studies on Cryptococcus mitochondrial inheritance and point out some important questions that need to be addressed in the future.

Plasma Circulating Cell-free Nuclear and Mitochondrial DNA as Potential Biomarkers in the Peripheral Blood of Breast Cancer Patients

  • Mahmoud, Enas H;Fawzy, Amal;Ahmad, Omar K;Ali, Amr M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8299-8305
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    • 2016
  • Background: In Egypt, breast cancer is estimated to be the most common cancer among females. It is also a leading cause of cancer-related mortality. Use of circulating cell-free DNA (ccf-DNA) as non-invasive biomarkers is a promising tool for diagnosis and follow-up of breast cancer (BC) patients. Objective: To assess the role of circulating cell free DNA (nuclear and mitochondrial) in diagnosing BC. Materials and Methods: Multiplex real time PCR was used to detect the level of ccf nuclear and mitochondrial DNA in the peripheral blood of 50 breast cancer patients together with 30 patients with benign lesions and 20 healthy controls. Laboratory investigations, histopathological staging and receptor studies were carried out for the cancer group. Receiver operating characteristic curves were used to evaluate the performance of ccf-nDNA and mtDNA. Results: The levels of both nDNA and mtDNA in the cancer group were significantly higher in comparison to the benign and the healthy control group. There was a statistically significant association between nDNA and mtDNA levels and well established prognostic parameters; namely, histological grade, tumour stage, lymph node status andhormonal receptor status. Conclusions: Our data suggests that nuclear and mitochondrial ccf-DNA may be used as non-invasive biomarkers in BC.

Morphological Variation and Partial Mitochondrial Sequence Analysis of Echinoid Species from the Coasts of the East Sea (동해 연안에 서식하는 성게의 형태변이와 미토콘드리아 유전자 분석)

  • Shin, Ji-Hye;Kim, Sung-Gyu;Kim, Young-Dae;Sohn, Young-Chang
    • Journal of Aquaculture
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    • v.21 no.3
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    • pp.139-145
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    • 2008
  • Morphological classification of echinoid species has many difficulties because of their phenotypic variations. In the present study, we analyzed morphotypes and partial mitochondrial 12S rDNA sequences of four sea urchin species classified as Pseudocentrotus depressus, Anthocidaris crassispina, Hemicentrotus pulcherrimus and Strongylocentrotus nudus, and unidentified four species collected from the coasts of the East sea. Their genomic DNAs were extracted from gonads and mitochondrial 12S rDNA sequences were amplified by the polymerase chain reaction (PCR) method. The sequence identities among the known four sea urchin species were 87.4-95.6%. The sequence identities among the unidentified four species were 99.4-99.6% and showed the highest homology to S. intermedius(99.8%). Thus, our phylogenetic tree indicates that the unidentified four species belong to S. intermedius.