• Journal of Life Science
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• v.19 no.4
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• pp.467-471
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• 2009

The entire D-loop region of the porcine mitochondrial DNA (mtDNA) was amplified from six pig breeds (Landrace, Duroc, Large White, Korean native pig, Berkshire, and Hampshire) using a primer set designed on the basis of reported porcine mtDNA sequences. From analyses through cloning, DNA sequencing and multiple sequence alignment, an 11-bp (TAAAACACTTA) duplication was observed after known tandem repetition in the D-loop region, which promoted hetroplasmy in mtDNA. Although the existence of the 11-bp duplication has been previously reported in Duroc and Japanese native pigs, there have not been any attempts to know the characteristics of this duplication in other breeds so far. A 150 bp fragment containing the 11-duplication was amplified and typed by polyacrylamide gel electrophoresis (PAGE). All Large Whites had two duplication units and Duroc showed heteromorphic patterns, 11.2% (9/80) of the animals had the 11-bp duplication in total. On the other hand, Landrace, Berkshire, Hampshire and Korean native pigs were non-duplicated. This result showed that the 11-bp duplication could be used as a breed-specific DNA marker for distinguishing pure Landrace and Large White breeds.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.3
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• pp.203-206
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• 2003

Objectives: Controversial arguments exists on both the case for and against on the accumulation of mitochondrial DNA (mtDNA) deletion in association to tissue and age. The debate continues as to whether this mutation is a major contributor to the phenotypic expression of aging and common degenerative diseases or simply a clinical insignificant epiphenomenon. The objective of this study was to determine whether the accumulation of mtDNA deletion is correlated with age-related and tissue-specific variation. Materials and Methods: One hundred and fifty-seven tissues from blood, ovary, uterine muscle, and abdominal muscle were obtained from patients ranging in age from 31$\sim$60 years. After reviewing the clinical reports, patients with mitochondrial disorder were excluded from this study. The tissues were obtained at gynecological surgeries with the consent of the patient. Total DNA isolated from blood, ovary, uterine muscle, and abdominal muscle was amplified by two rounds of PCR using two pairs of primers corresponding to positions 8225-8247 (sense), 13551-13574 (antisense) for the area around deleted mtDNA and 8421-8440 (sense), 13520-13501 (antisense) for nested PCR product. A statistical analysis was performed by $x^2$-test. Results: About 0% of blood, 94.8% of ovary, 71.4% of uterine muscle, and 86.1% abdominal muscle harbored mtDNA deletion. When we examined the proportion of deleted mtDNA according to age deletion rate was 90% of ovary, 63.6% of uterine muscle, 77.7% of abdominal muscle in thirties and 100% of all tissue in fifties. Conclusion: The findings of this study suggest that the mtDNA deletion is varied in tissue-specific pattern and increases with aging.

• Journal of Nutrition and Health
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• v.48 no.5
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• pp.390-397
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• 2015

Purpose: In this study, we investigated the effects of chronic alcohol and excessive iron intake on mitochondrial DNA (mtDNA) damage and the progression of alcoholic liver injury in rats. Methods: Twenty-four Sprague-Dawley male rats were divided into four groups (Control, EtOH, Fe, and EtOH + Fe), and fed either control or ethanol (36% of total calories) liquid diet with or without 0.6% carbonyl iron for eight weeks. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, liver malondialdehyde concentrations were measured by colorimetric assays. Liver histopathology was examined by Hematoxylin-eosin staining of the fixed liver tissues. The integrity of the hepatic mtDNA and nuclear DNA was measured by long-range PCR. The gene expression levels of cytochrome c oxidase subunit 1 (Cox1) and NADH dehydrogenase subunit 4 (Nd4) were examined by real-time PCR. Results: Serum ALT and AST activities were significantly higher in the EtOH+Fe group, as compared to the Control group. Similarly, among four groups, liver histology showed the most severe lipid accumulation, inflammation, and necrosis in the EtOH + Fe group. PCR amplification of near-full-length (15.9 kb) mtDNA showed more than 50% loss of full-length product in the liver of the EtOH + Fe group, whereas amounts of PCR products of a nuclear DNA were unaffected. In addition, the changes in the mtDNA integrity showed correlation with reductions in the mRNA levels of mitochondrial gene Cox1 and Nd4. Conclusion: Our data suggested that the liver injury associated with excessive iron and alcohol intake involved mtDNA damage and corresponding mitochondrial dysfunction.

• Analytical Science and Technology
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• v.18 no.3
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• pp.257-262
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• 2005

Mitochondrial DNA (mt DNA) sequence analysis has been a useful tool for species identification of animals and human individuals. Two hypervariable regions (HV1 and HV2) in control region of mitochondrial genome were analyzed for human individual identification. In case of animal species identification, several genes on mt DNA such as cytochrome b (cytb), RNAs, cytochrome oxidases (CO) were used. In this study, co-amplification of HV1 and cytb was carried out in order to check the contamination of animal DNA and to verify the human DNA. The primer sets used in PCR were H15997/L16236 for HV1 and H14724/L15149 for cytb. PCR products for HV1 and cytb were 239 bp and 425 bp, respectively. The appearance of two bands on agarose gel implied the DNA came from human, however the single band of cytb gene represented the non-human animal DNA.

• Animal Systematics, Evolution and Diversity
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• v.36 no.2
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• pp.182-184
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• 2020

Isaacsicalanus paucisetus Fleminger, 1983, a monotypic species of the family Spinocalanidae Vervoort, 1951, was first reported from a hydrothermal vent field in the East Pacific Rise off the mouth of the Gulf of California. The mitochondrial cytochrome oxidase I(mtCOI) DNA barcodes are considered a useful tool to assist traditional taxonomy and species discrimination in calanoid copepods. However, the mtCOI DNA barcodes of I. paucisetus have not been reported due to the species rarity and the difficulty of sampling. In this study, we firstly determined the mtCOI DNA barcodes of the I. paucisetus newly collected from a hydrothermal vent in the North Fiji Basin of the southwestern Pacific. All mtCOI DNA barcodes of I. paucisetus were identical and intraspecies variations of spinocalanid species were 0.0-3.0%. Interspecies and intergeneric variations were 13.4-25.2% and 16.7-24.1%, respectively. The DNA barcodes of I. paucisetus obtained in the present study would be helpful for understanding taxonomic relationships of widespread spinocalanid species.

• Animal cells and systems
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• v.3 no.1
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• pp.79-87
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• 1999

Mitochondrial DNA (mtDNA) from 54 specimens of the blue mussel (Mytilus edulis) species complex sampled from the southern coast of Korea was assayed for polymorphism with a portion of the COIII gene (336 bp). Fifteen haplotypes were found. PAUP, one-step networks, and PHYLIP analyses revealed the presence of two clearly differentiated mitochondrial clades (termed clades B and E), separated by 3.6% of minimum sequence divergence. The distribution pattern of the species appears to be consistent with category II of the phylogeographic pattern sensu (Avise et al., 1987): the presence of two discontinuous and distinct mtDNA genotypes in the same geographic region. This unusual mitochondrial polymorphism was explained by the presence of the Mediterranean species, M. galloprovincialis, possessing mtDNA of both M. galloprovincialis and M. edulis.

• The Korean Journal of Zoology
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• v.37 no.3
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• pp.439-451
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• 1994

미꾸리속 어류 2종의 유전적 차이를 알아보기 위하여 염색체 분석과 미토콘드리아 DNA(mtDNA) 분석을 실시하였다. 일반염색에 의한 핵형 분석 결과 미꾸리(2N=50)와 미꾸라지(2N=48)는 염색체수에 차이가 있었으며. N-banding 분석 결과 두 종간에는 인형성 부위의 위치와 크기에 차이가 있었다. C-handing 겪과 미꾸라지는 1번 염색체쌍 동원 체부위에 넓게 C-band플 갖고 있었다. 미꾸리속 어류 2종의 mtONA를 7개의 6-base cutting 제한효소로 처리하여 절편 양상을 비교. 분석한 결과 2종 공히 mtDNA의 genome 크기는 약 16.OKb였으며 fragment homology(F)에서 미꾸리의 종내 집단간의 F값은 0.674. 미꾸라지는 0.862로 유사하게 나타났으나, 종간 F값은 0.207(0.074-0.417)로 낮았다 염기치환율은 미꾸리가 p=0.021, 미꾸라지는 p=0.002로 미꾸라지가 미꾸리에 비해 매우 낮은 염기치환율을 보였고. 종간 평균 염기치환율은 p=0.104로 차이 를 나타냈다. MtDNA 분석과 핵형 분석 결과 미꾸라지는 Robertsonlan translocation의 결과 미꾸리로 부터 분화된 것으로 추정 되었다.

• Journal of Animal Science and Technology
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• v.46 no.6
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• pp.917-924
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• 2004

Using PCR-RFLP haplotyping for the mitochondrial DNA(mtDNA) fragment containing the NADH dehydrogenase 2 gene(ND2) and three tRNA genes(tRNA-Met, tRNA-Trp and tRNA-Ala), we characterized the genetic diversity of five pig breeds including Jeju native pigs. mtDNA polymorphisms showing distinct cleavage patterns were found in the pig breeds. Two digestion patterns were detected when HaeIII- and Hinfl-RFLP, and four in the Tsp5091-RFLP analyses. Combining the three restriction enzyme digestion patterns found in five different pig breeds, four mtDNA haplotypes were observed and the haplotype frequencies were significantly different by the pig breeds. A monomorphic haplotype, mtWB, was observed in both Korean wild boars and Large White pigs. Both Duroc and Landrace pigs contained two haplotypes suggesting their multiple maternal lineages. Jeju native pig has two haplotypes(mtJN and mtJD). Of these, mtJN is identified as a Jeju native pig specific haplotype. This study suggested that more than two progenitor populations have been taken part in the domestication process of the Jeju native pig population, and/or probably subsequent crossing with other pig breeds from near east Asia. Unlike with our prediction, there was no direct evidence under molecular levels on the maternal introgression of Korean wild boar in the domestication of Jeju native pigs. In conclusion, specificity of mtDNA haplotypes related to pig breeds win be useful for identifying the maternal lineage as wen as constructing the genealogical pedigree in pigs.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.6
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• pp.474-484
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• 1992

The stock identification of small yellow croaker. Pseudosciaena Polyactis from Mokpo area was carried out using molecular biological methods such as mt-DNA restriction fragment length polymorphism(RFLP) and the N-terminal fragment polymorphism of muscle actin obtained after protease digestion. The entire mt-DNA genomic size from the small yellow croaker at Mokpo area was estimated to be about $16\pm0.2$ Kb. Furthermore, fourteen restriction endonucleases revealed a total of 37 restriction sites to the mt-DNA molecule, however, eight of the fourteen enzymes showed a significant restriction site variation. Six of the enzymes examined produced a single restriction profile for all individuals surveyed, indicating that they don't react on the same mt-DNA obtained from small yellow croaker. The Staphylococcus aureus $V_8$ protease is able to cleave the muscle actin of small yellow croaker and to yield a N-terminal peptide of 26 and 16 KDa, respectively.

• Journal of Life Science
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• v.14 no.6 s.67
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• pp.913-919
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• 2004

Mitochondrial DNA mutations have been reported in recent years in association with sensorineural hering loss. The purpose of this study is to identify the association between the noise-induced sensorineural hearing loss and the A to G mutation at nucleotide 3243, 1555, 7445 of mitochondrial DNA. Study subjects were established by history and chart review, and audiological and clinical data were obtained. Blood was sampled from 214 normal controls, 102 noise-induced hearing loss, and 28 sensorineural hearing loss. The DNA of these individuals were extracted, and mitochondrial DNA fragments were analyzed by polymerase chain reaction. Subsequently, the coding sequence of mitochondrial DNA 3243, 1555, 7445 were sequenced, and compared to the normal sequence, and all sequence variations were analyzed by restriction enzymes. Mitochondrial DNA mutations $(3243A{\rightarrow}G,\;1555A{\rightarrow}4G,\;7445A{\rightarrow}G)$ were not detected by polymerase chain reactions in any patients with noise-induced hearing loss, sensorineural hearing loss, and normal controls. The DNA sequencing of PCR products did not revealed an A to G substitution at nucleotide 3243, 1555, 7445 of mitochondrial DNA. The noise-induced sensorineural hearing loss was not associated with mitochondrial DNA mutation $(3243A{\rightarrow}G,\;1555A{\rightarrow}4G,\;7445A{\rightarrow}G)$.