• Title/Summary/Keyword: mitochondrial marker

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An Introduction to Microsatellite Development and Analysis (Microsatellite 개발 및 분석법에 대한 소개)

  • Yun Young-Eun;Yu Jeong-Nam;Lee Byoung-Yoon;Kwak Myounghai
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.299-314
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    • 2011
  • The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.

Application of Breed-specific DNA Markers for the use of Identifying Major Pure Pig Breeds Maintained in Korea (대한민국내 주요 돼지 품종의 순종 식별을 위한 품종특이 DNA marker의 활용)

  • Seo, B.Y.;Kim, J.H.;Park, E.W.;Lim, H.T.;Cho, I.C.;Kim, B.W.;Oh, S.J.;Cheong, I.C.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.735-742
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    • 2004
  • This study was conducted for the identification of pure Landrace, Large White and Duroc breeds which are mainly maintained in Korea using DNA markers. We used known KIT and MC1R mutations, which were related coat color in pigs, and pig mitochondrial DNA variations. The KIT mutation was used to distinguish white and colored animals. Duroc breed could be discriminated from other colored breeds using the MC1R mutation N121D. Discriminating Landrace and Large White was possible using the l l-bp duplication of D-Ioop region and alternative initiation codon of ND2. In conclusion, identification of Landrace, Large White and Duroc breeds was might be possible using the procedure designed in this study.

The Effects of Isocaloric High-Fat Diet and Endurance Exercise on Insulin Resistance and Mitochondria Biogenesis (Isocaloric high-fat diet와 지구성 운동이 인슐린 저항성과 미토콘드리아 생합성에 미치는 영향)

  • Jung, Su-Ryun;Kwon, Dae-Yong;Kim, Jin-Hwan;Kang, Ho-Youl
    • 한국체육학회지인문사회과학편
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    • v.51 no.3
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    • pp.353-362
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    • 2012
  • The purpose of this research was to find effects of high fat diet and endurance exercise on mitochondria biosynthesis and insulin resistance in male Wister rat. Th rats were randomly separated into 4 groups: A, Isocaloric high fat diet group (50% of calories from fat), B. Chow group, C. Isocaloric high fat diet with endurance exercise (treadmill running, slop 8%, 23 m/min, 120 min/day, 5 days per week), D. Chow group with endurance exercise. Both Isocaloric high fat diet group and chow group were given an equal caloric composition with 3.2kcal/g. equal amount of food were checked every day and given to both groups. 4 weeks of high fat diet did not show any change in body weight and amount of body fat. Further the level of glucose and insulin in blood, and insulin-stimuilated glucose transport rate in epitrochlearis muscle was not affected by Isocaloric high fat diet. However, the endurance exercise showed statistically significant change in the level of insulin in blood. Although either Isocaloric high fat diet or endurance exercise alone did not change on mitochondria biogenesis marker, Isocaloric high fat diet with endurance exercise could induce the increased level of marker (p<0.05). Also, plasma free fatty acids were increased in this group. From this investigation, Isocaloric high fat diet with moderate-intensity endurance exercise is effective way to induce mitochondrial biogenesis.

Species-specific Marker Development for Environmental DNA Assay of Endangered Bull-head Torrent Catfish, Liobagrus obesus (멸종위기어류 퉁사리의 환경 DNA 분석을 위한 종 특이 마커 개발)

  • Yun, Bong Han;Kim, Yong Hwi;Sung, Mu Sung;Han, Ho-Seop;Han, Jeong-Ho;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.34 no.3
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    • pp.208-217
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    • 2022
  • We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.

Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea

  • Won, Sohyun;Park, Bae Keun;Kim, Baek Jun;Kim, Hye Won;Kang, Jun Gu;Park, Tae Seo;Seo, Hong Yul;Eun, Ye;Kim, Ki Gyoung;Chae, Joon Seok
    • Parasites, Hosts and Diseases
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    • v.52 no.2
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    • pp.169-175
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    • 2014
  • There are 60 species of blood-feeding land leeches, 50 species belonging to the family Haemadipsidae and 10 species belonging to the family Xerobdellidae. Despite recent papers on the land leeches, their taxonomic identification is not fully understood, especially at a species level. In Korea, there have been no historical records of the terrestrial leeches, but recently an unrecorded blood-feeding land leech was discovered at Gageo-do (Island), Korea. Molecular analysis was used to identify the species of 29 leeches collected from Mt. Dock-Sil in Gageo-do. Conventional PCR was conducted using nuclear 18S rRNA and mitochondrial cytochrome c oxidase subunit 1 (CO1) genetic marker. The 18S rRNA sequences revealed that the leeches share 99.9% identity with Haemadipsa rjukjuana (inhabiting Taiwan), and the CO1 sequences revealed that the leeches are very close to H. rjukjuana (inhabiting Taiwan). The CO1 sequences were separated into 2 categories, 1 with 94.6% and the other with 94.3% similarity to the H. rjukjuana L00115A (inhabiting Taiwan). This new finding of the land leech is the first record in Korea. In addition, the north range of the distribution of the blood-feeding leech (Hirudiniformes: Haemadipisidae) should be reconsidered including Korea.

Regulation of PPAR and SREBP-1C Through Exercise in White Adipose Tissue of Female C57BL/6J Mice

  • Jeong, Sun-Hyo
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.227-236
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    • 2012
  • Previous study showed that swimming improved obesity but was not through $PPAR{\alpha}$ activation in liver and skeletal muscle in high fat diet-fed female mice with functioning ovaries as an animal model of obese premenopausal women. Thus, this study was aimed at investigation of the effects of swimming on the promotion of health and its molecular mechanism in adipose tissue of high fat diet-fed female mice. Eight-week-old female C57BL/6J mice were randomly divided into two groups (a non-swim control group and a swim group, n=8/group). Mice in the swim group swam for 2 h daily for 6 weeks in water bath with temperature of $35{\pm}1^{\circ}C$. All the animals received high fat diet (45% kcal fat) for 6 weeks. Reverse transcription-polymerase chain reaction was used to elucidate the molecular mechanism. Female mice subjected to swimming had significantly decreased body weight gain and white adipose tissue mass compared with the female control mice. Histological studies illustrated that swimming decreases the hepatic lipid accumulation. As expected, swimming did not affect the expression of mRNA levels of peroxisome proliferator-activated receptor (PPAR) ${\alpha}$ and $PPAR{\alpha}$ target genes responsible for mitochondrial fatty acid ${\beta}$-oxidation, such as carnitine palmitoyltransgerase-1 and medium chain acyl-CoA dehydrogenase in the white adipose tissue. However, mice that underwent 6-weeks of swimming exercise had decreased the mRNA expression of lipogenic genes, such as sterol regulatory element-binding proteins-1C and fatty acid synthase in comparison to sedentary control mice, with decreased $PPAR{\gamma}$ target genes involved in adipocyte-specific marker genes, such as adipocyte fatty acid binding protein and leptin in the white adipose tissue. These results suggest that swimming can effectively prevent obesity induced by high fat diet-fed, in part through down-regulation of adipogenesis and lipogenesis in white adipose tissue of female obese mice. Moreover, these results suggest that swimming maybe contributing the promotion of health through regulation of adipogenesis and lipogenesis in overweight premenopausal women.

Fuctional Relationship between Rate of Fatty Acid Oxidation and Carnitine Palmitoyl Transferase I Activity in Various Rat Tissues

  • Cho, Yu-Lee;Do, Kyung-Oh;Kwon, Tae-Dong;Jang, Eung-Chan;Lee, Keun-Mi;Lee, Suck-Kang;Kim, Jong-Yeon
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.4
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    • pp.207-210
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    • 2003
  • Lipids play many structural and metabolic roles, and dietary fat has great impact on metabolism and health. Fatty acid oxidation rate is dependent on tissue types. However there has been no report on the relationship between the rate of fatty acid oxidation and carnitine transport system in outer mitochondrial membrane of many tissues. In this study, the rate of fatty acid oxidation and carnitine palmitoyltransferase (CPT) I activity in the carnitine transport system were measured to understand the metabolic characteristics of fatty acid in various tissues. Palmitic acid oxidation rate and CPT I activity in various tissues were measured. Tissues were obtained from the white and red skeletal muscles, heart, liver, kidney and brain of rats. The highest lipid oxidation rate was demonstrated in the cardiac muscle, and the lowest oxidation rate was in brain. Red gastrocnemius muscle followed to the cardiac muscle. Lipid oxidation rates of kidney, white gastrocnemius muscle and liver were similar, ranging from 101 to 126 DPM/mg/hr. CPT I activity in the cardiac muscle was the highest, red gastrocnemius muscle followed by liver. Brain tissue showed the lowest CPT I activity as well as lipid oxidation rate, although the values were not significantly different from those of kidney and white gastrocnemius muscle. Therefore, lipid oxidation rate was highly (p<0.001) related to CPT I activity. Lipid oxidation rate is variable, depending on tissue types, and is highly (p<0.001) related to CPT I activity. CPT I activity may be a good marker to indicate lipid oxidation capacity in various tissues.

Development of Molecular Marker to Distinguish Octopus minor Sasaki Caught in Korea and that in China (한국산과 중국산 낙지구별을 위한 DNA 마커)

  • Kim, Joo-Il;Oh, Taeg-Yun;Yang, Won-Seok;Cho, Eun-Seob
    • Journal of Life Science
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    • v.18 no.2
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    • pp.284-286
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    • 2008
  • Octopus minor (O. minor) is widely distributed along the coastal regions of Korea, but most of them are caught in southern waters which are associated with one of the important fisheries stock. At present, O. minor from China has been introduced to the fishery markets in Korea. Here, we attempt to discriminate their origin for Korea or China using molecular techniques. Based on the O. minor mitochondrial DNA sequence, we developed a PCR-based origin discrimination system. The assay specificity was assessed by testing four individuals of O. minor from Sangdong, China, as well as 20 additional O. minor from Namhae, Muan, Yeosu and Jindo, Korea. Only four isolates of O. minor originated from China tested as positive in our distinction system. All PCR-positive products yielded identical sequences from Chinese O. minor, whereas Korean O. minor appeared to be PCR amplification. This result suggested that the primers used in this study are O. minor species specific, especially originated from China. The detection system appeared to be positive results in the use of 0.1 ng of Chinese O. minor DNA as template, however, the Korean O. minor even using $1{\mu}g$ of DNA showed no amplification. Consequently, the assay provides a simple, rapid and accurate method for the detection of Chinese O. minor.

Effects of Dimethyl Sulfoxide on the Differentiation of Myocardial and Endothelial Cells (심근세포 및 내피새포의 분화에 미치는 Dimethyl Sulfoxide의 영향)

  • Lee, Dong-Hyup;Park, Yee-Tae;Han, Sung-Sae;Lee, Yung-Chang
    • Journal of Yeungnam Medical Science
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    • v.5 no.2
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    • pp.111-119
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    • 1988
  • To elucidate the effects of dimethyl sulfoxide on myocardial and endothelial cells in culture, the cells were exposed to 10% dimethyl sulfoxide in culture medium for 1 hour at 48 hours after cell isolation. The general morphology and the cytochemical reaction of marker enzymes for mitochondria and Golgi complexes were investigated. The results were summarized as follows. : 1. DMSO induced elongation and narrowing of the cells and increase of mitochondrial reaction in myocardial cells. 2. DMSO induced destruction and disruption of myofibrils in myocardial cells resulting in increase of contractile activities. 3. In the endothelial cells, DMSO suppressed proliferative activities but thiamine pyrophosphatase reactions were enhanced indicating increase of Goigi complex activity. 4. DMSO seemed to hamper with the adhesiveness and motility of the endothelial cells causing the decrease of the number of cells in vitro.

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The rs196952262 Polymorphism of the AGPAT5 Gene is Associated with Meat Quality in Berkshire Pigs

  • Park, Woo Bum;An, Sang Mi;Yu, Go Eun;Kwon, Seulgi;Hwang, Jung Hye;Park, Da Hye;Kang, Deok Gyeong;Kim, Tae Wan;Park, Hwa Chun;Ha, Jeongim;Kim, Chul Wook
    • Food Science of Animal Resources
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    • v.37 no.6
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    • pp.926-930
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    • 2017
  • High-quality meat is of great economic importance to the pig industry. The 1-acylglycerol-3-phosphate-O-acyltransferase 5 (AGPAT5) enzyme converts lysophosphatidic acid to phosphatidic acid in the mitochondrial membrane. In this study, we found that the porcine AGPAT5 gene was highly expressed in muscle tissue, influencing meat characteristics, and we also identified a non-synonymous single-nucleotide polymorphism (nsSNP) (rs196952262, c.673 A>G) in the gene, associated with a change of isoleucine 225 to valine. The presence of this nsSNP was significantly associated with meat color (lightness), lower cooking loss, and lower carcass temperatures 1, 4, and 12 h after slaughter (items T1, T4, and T12 on the recognized quality scale, respectively), and tended to increase backfat thickness and the water-holding capacity. These results suggest that nsSNP (c.673A>G) of the AGPAT5 gene is a potential genetic marker of high meat quality in pigs.