• Title/Summary/Keyword: mitochondrial gene

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Phylogenetic Analysis Using Cytochrome c Oxidase Subunit I of Silver Croaker(Pennahia argentata) Mitochondria DNA (미토콘드리아 DNA의 cytochrome c oxidase subunit I을 이용한 보구치(Pennahia argentata) 계통 분석)

  • Park, Jae-Won;Park, Kiyun;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.53 no.3
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    • pp.265-274
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    • 2020
  • Silver croaker (Pennahia argentata) is a turbulent species that is widely distributed worldwide and is mainly found in the bottom of the ocean. In the study, we characterized the cytochrome c oxidase subunit I (COI) gene of the mitochondrial DNA (mtDNA) on P. argentata inhabiting Gwangyang Bay and analyzed the phylogenetic location of marine fish species. As a result of multiple arrangement of 605 bp COI sequences, high homology of mtDNA nucleotide sequences was confirmed in the silver croakers from Gwangyang Bay (98~100%). However, the nucleotide variation was different according to the catching points of the inland and the open seas of Gwangyang Bay. The nucleotide sequence variation in COI was high in P. argentata from the open seas of Gwangyang Bay (43.2~70.3%). Furthermore, the phylogenetic analysis of 13 fish showed that P. argentata from Gwangyang Bay were grouped into one clade with P. argentata reported in Taiwan, and the evolutionary distance was 0.036. In addition, it was identified that the evolutionary distance was close to that of fish belonging to the Mi-iuy croaker (Miichthys miiuy) and the Big-head pennah croaker (Pennahia Macrocephalus) (0.041~0.048). The result of these studies will be used as the key genetic information for fisheries resources monitoring and species diversity management according to the coastal environment.

Clinical Features, Response to Treatment, Prognosis, and Molecular Characterization in Korean Patients with Inherited Urea Cycle Defects

  • Yoo, Han-Wook;Kim, Gu-Hwan;Seo, Eul-Ju
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.2 no.1
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    • pp.77-79
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    • 2002
  • The urea cycle, consisting of a series of six enzymatic reactions, plays key roles to prevent the accumulation of toxic nitrogenous compound and synthesize arginine de novo. Five well characterized diseases have been described, resulting from an enzymatic defect in the biosynthesis of one of the normally expressed enzyme. This presentation will focus on two representative diseases; ornithine transcarbamylase(OTC) deficiency and citrullinemia(argininosuccinate synthetase deficiency). OTC deficiency is one of the most common inborn error of urea cycle, which is inherited in X-linked manner. We identified 17 different mutations in 20 unrelated Korean patients with OTC deficiency; L9X, R26P, R26X, T44I, R92X, G100R, R141Q, G195R, M205T, H214Y, D249G, R277W, F281S, 853 del C, R320X, V323M and 10 bp del at nt. 796-805. These mutations occur at well conserved nucleotide sequences across species or CpG hot spot. The L9X and R26X lead to the disruption of leader sequences, required for directing mitochondrial localization of the OTC precursor. Their phenotypes are severe, and neonatal onset. The G100R, R277W and V323M mutations were uniquely identified in patients with late onset OTC deficiency. The other genotypes are associated with neonatal onset. Out of 20 patients with OTC deficiency, only 6 patients are alive; two were liver transplanted, and normal in growth and development at 2, 4 years after transplantation respectively. Citrullinemia is an autosomal recessive disease, caused by the mutations in the argininosuccinate synthetase(ASS) gene. We identified in 3 major mutations in 11 unrelated Korean patients with citrullinemia; G324S, $IVS6^{-2}$ A to G, and 67 bp ins at nt 1125-1126. Among these, the 67 base pair insertion mutation is novel. The allele frequency of each mutation is; G324S(45%), IVS6-2 A to G(32%), and 67 base pair insertion(14%). All patients are diagnosed at neonatal or infantile age. Interestingly, two patients presented with stroke like episode. Out of 11 patients, 5 patients died. Among 6 patients alive, one patient was successfully liver transplanted.

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Development of a Multiplex PCR Assay for Rapid Identification of Larimichthys polyactis, L. crocea, Atrobucca nibe, and Pseudotolithus elongates (다중 PCR 분석법을 이용한 참조기, 부세, 흑조기 및 긴가이석태의 신속한 종판별법 개발)

  • Noh, Eun Soo;Lee, Mi-Nan;Kim, Eun-Mi;Park, Jung Youn;Noh, Jae Koo;An, Cheul Min;Kang, Jung-Ha
    • Journal of Life Science
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    • v.27 no.7
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    • pp.746-753
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    • 2017
  • In order to rapidly identify four drums species, Larimichthys polyactis, L. crocea, Atrobucca nibe, and Pseudotolithus elongates, a highly efficient and quick method has been developed using multiplex polymerase chain reaction (PCR) with species-specific primers. Around 1.4 kbp of the mitochondrial COI gene sequences from the four drums species were aligned, and species-specific forward primers were designed, based on the single nucleotide polymorphism (SNP). The optimal conditions for PCR amplification were selected through cross-reactivity, using a gradient PCR method. The PCR results demonstrated species-specific amplification for each species at annealing temperatures between 50 and $62^{\circ}C$. Multiplex species-specific PCR (MSS-PCR) amplification reactions with four pairs of primers were performed for sixteen specimens of each species. MSS-PCR lead to a species-specific amplification of a 1,540 bp fragment in L. polyactis, 1,013 bp in A. nibe, 474 bp in L. crocea, and 182 bp in P. elongates, respectively. The four different sizes of each PCR product can be quickly and easily detected by single gel electrophoresis. The sensitivity of the MSS-PCR of the DNA was up to $0.1ng/{\mu}l$ as a starting concentration for the four different species tested. These results suggest that MSS-PCR, with species-specific primers based on SNP, can be a powerful tool in the rapid identification of the four drums species, L. polyactis, L. crocea, A. nibe, and P. elongates.

Mitochondrial Genetic Diversity and Phylogenetic Relationships of Siberian Flying Squirrel(Pteromys volans) Populations

  • Lee, Mu-Yeong;Park, Sun-Kyung;Hong, Yoon-Jee;Kim, Young-Jun;Voloshina, Inna;Myslenkov, Alexander;Saveljev, Alexander P.;Choi, Tae-Young;Piao, Ren-Zhu;An, Jung-Hwa;Lee, Mun-Han;Lee, Hang;Min, Mi-Sook
    • Animal cells and systems
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    • v.12 no.4
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    • pp.269-277
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    • 2008
  • Siberian flying squirrel, an endangered species in South Korea, is distributed through major mountain regions of South Korea. The number of Siberian flying squirrel(Pteromys volans) in South Korea has decreased and their habitats are fragmented and isolated because of anthropogenic activities. So far no molecular genetic data has, however, been available for their conservation and management. To obtain better information concerning genetic diversity and phylogenetic relationships of the Siberian flying squirrel in South Korea, we examined 14 individuals from South Korea, 7 individuals from Russia, and 5 individuals from northeastern China along with previously published 29 haplotypes for 1,140 bp of the mtDNA cytochrome b gene. The 14 new individuals from South Korea had 7 haplotypes which were not observed in the regions of Russia and Hokkaido. The level of genetic diversity(0.616%) in the South Korean population was lower than that in eastern Russia(0.950%). The geographical distribution of mtDNA haplotypes and reduced median network confirmed that there are three major lineages of Siberian flying squirrel, occupying; Far Eastern, northern Eurasia, and the island of Hokkaido. The South Korean population only slightly distinct from the Eurasia, and eastern Russian population, and is part of the lineage Far Eastern. Based on these, we suggest that the South Korean population could be considered to belong to one partial ESU(Far Eastern) of three partial ESUs but a different management unit. However, the conservation priorities should be reconfirmed by nuclear genetic marker and ecological data.

Development of TaqMan probe-based real-time PCR for rapid identification of beef, pork and poultry meat (소, 돼지, 가금육류의 신속한 동정을 위한 TaqMan probe를 이용한 real-time PCR 개발)

  • Koh, Ba-Ra-Da;Kim, Ji-Yeon;Na, Ho-Myung;Park, Seong-Do;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.215-222
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    • 2012
  • Species-specific $TaqMan^{(R)}$ probe-based real-time PCR assays were developed for detection of beef, pork, chicken, duck, goose and turkey. The primer and probe sets used in this study were designed to be complementary to fibroblast growth factor (FGF) for cattle and pig, mitochondrial NADH dehydrogenase (ND) subunit 3 and ND2 for chicken and duck, 12S rRNA for goose and turkey, respectively. As internal positive control we used conserved region in the ribosomal 18S RNA gene to ensure the accuracy of the detection of target DNA by real-time PCR. We confirmed that real-time PCR assays with the primer and probe sets were positive for cattle, pig and chicken intended target animal species with no cross-reactivity with other non-target animal species. Only >50 ng DNA of beef show cross-reactivity in the determination of duck. Using species-specific primer and probe sets, it was possible to detect amounts of 0.1 ng DNA of cattle and pig, 1.0 pg DNA of chicken, duck and turkey, and 0.1 pg DNA of goose for raw samples, respectively. The detection limits were 0.1 ng DNA of cattle, 1.0 ng DNA of pig and 1.0 pg DNA of chicken for DNA mixtures (beef, pork and chicken) extracted from heat-treated ($121^{\circ}C$/5 min) meat samples. In conclusion, it can be suggested that the $TaqMan^{(R)}$ probe-based assay developed in this study might be a rapid and specific method for the identification of meat species in raw or cooked meat products.

Rapid and Specific Identification of Genus Cynoglossus by Multiplex PCR Assays Using Species-specific Derived from the COI Region (다중 PCR 분석법을 이용한 참서대과 어종의 신속하고 정확한 종판별 분석법 개발)

  • Noh, Eun Soo;Kang, Hyun Sook;An, Cheul Min;Park, Jung Youn;Kim, Eun Mi;Kang, Jung Ha
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1007-1014
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    • 2016
  • A highly efficient, rapid, and reliable multiplex polymerase chain reaction based method for distinguishing ten species of genus Cynoglossus (C. senegalensis, C. abbreviates, C. macrolepidotus, C. arel, C. semilaevis, C. interruptus, C. joyneri, C. lingua, C. robustus, and C. monodi) is described. The species-specific primer sets were designed base on the cytochrome oxidase subunit I gene (1,500 bp). The optimal PCR conditions and primers were selected for ten of Cynoglossus species to determine target base sequences using single PCR. Multiplex PCR using the ten pairs of primers either specifically amplified a DNA fragment of a unique size or failed, depending on each species DNA. The length of amplification fragment of 208 bp for C. senegalensis, 322 bp for C. abbreviates, 493 bp for C. macrolepidotus, 754 bp for C. arel, 874 bp for C. semilaevis, 952 bp for C. interruptus, 1,084 bp for C. joyneri, 1,198 bp for C. lingua, 1,307 bp for C. robustus, and 1,483 bp for C. monodi with the species-specific primers, visualized by agarose gel electrophoresis, allowed perfectly distinction of the Cynoglossus species. The multiplex PCR assay can be easily performed on multiple samples and attain final results in less than 6 hours. This technique should be a useful addition to the molecular typing tools for the tentative identification of Cynoglossus species.

Molecular Identification and Distribution Pattern of Fish Eggs Collected around Jejudo Island (제주 연안해역 어란의 분자동정과 분포양상)

  • Han, Song-Hun;Kim, Maeng Jin;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.27 no.4
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    • pp.284-292
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    • 2015
  • Monthly variation of species composition and abundance of fish eggs were examined to know the spawning time and location of the fishes inhabiting the coastal region of Jejudo Island. Samplings had been performed at the four locations (Jeju port, Seongsanpo, Seogwipo port and Chagwido) with a bongo net which was towed monthly at the sea surface from August 2006 to July 2007. The fish eggs were identified based on phylogenetic analyses with the DNA sequences generated through PCR-amplification and sequencing of the mitochondrial cytochrome b gene. Among a total of 43 taxa classified during the study period, 34 were identified to species, 4 to families, and the remaining 5 were unidentified. Of them, 23 taxa were occurred at Jeju port, 21 at Seongsanpo, 19 at Seogwipo port and 18 at Chagwido, whereas 15 taxa were found in September 2006, 12 in June 2007, 6 to 8 in August 2006 and January~May 2007, 5 in each October and November 2006, 3 in each December 2006 and July 2007. Among 34 species of fish eggs, Engraulis japonicus and Callanthias japonicus most frequently appeared at 16 times out of 48 observations in total. When those eggs were collected, the surface seawater temperature ranged $14.0{\sim}28.6^{\circ}C$ for E. japonicus and $14.9{\sim}20.5^{\circ}C$ for C. japonicus. The success rates of PCR-amplification and species identification in this study were 68.3% and 79.1%, respectively.

mtDNA Diversity and Phylogenetic Analysis of Korean Native Goats (한국재래염소의 mtDNA 다양성 및 계통유전학적 분석)

  • Kim, Jae-Hwan;Cho, Chang-Yeon;Choi, Seong-Bok;Cho, Young-Moo;Yeon, Seung-Hum;Yang, Boh-Suk
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1329-1335
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    • 2011
  • Korean native goats, which are characterized by black coat color, have existed on the Korean peninsula for a long time. Until now, there has been no comprehensive investigation concerning their genetic diversity, phylogenetic analysis or origin. In this study, we investigated the genetic diversity and verified phylogenetic status of the Korean native goat using the 453-bp fragment of the hypervariable fragment I (HVI) of mitochondrial DNA (mtDNA) D-loop region from 60 individuals among 5 populations. The Korean native goat showed less haplotype diversity when compared with goats from other countries. In addition, 6 haplotypes that had not been previously reported were verified in this study. In phylogenetic analyses with other country's goats, 10 haplotypes from Korean native goats were classified into mtDNA lineage A. Moreover, in a phylogenetic tree for goats which contained mtDNA lineage A, 8 of 10 haplotypes could be included in a subgroup with goats from Vietnam and an area of China. However, none of the remaining haplotypes belonged to a major group of Korean native goats and were located on different independent positions. These results suggest that almost Korean native goats aligned more closely to China and Vietnam breeds in mtDNA lineage A and there was no gene flow from other mtDNA lineages. Our results will contribute to conservation strategies and genetic breeding of Korean native goats.

Genetic Population Structure and Phylogenetic Relationship of the Large-footed Bat (Myotis macrodactylus) on Jeju Island (제주도 큰발윗수염박쥐(Myotis macrodactylus)의 유전적 집단 구조와 계통 유연관계)

  • Kim, Yoo-Kyung;Park, Su-Gon;Han, Sang-Hoon;Han, Sang-Hyun;Oh, Hong-Shik
    • Journal of Life Science
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    • v.26 no.7
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    • pp.749-757
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    • 2016
  • This study was carried out to reveal the genetic population structure of the Jeju Island population and the phylogenetic relationship of East Asian populations of the large-footed bat (Myotis macrodactylus) based on the genetic polymorphisms of mitochondrial cytochrome B (CYTB) and NADH dehydrogenase subunit 1 (ND1) gene sequences. A total of fourteen and nine haplotypes were found in the CYTB and ND1 sequences from East Asian bats, respectively. Haplotype distribution showed locality specific patterns. The results from ND1 haplotype analysis showed that the Jeju Island population has four haplotypes: the Mt. Halla and Western subpopulations have three ND1 haplotypes, but the Eastern subpopulation has just a single haplotype Nd03, which is commonly found on this island. The neighbor-joining (NJ) tree showed the closer relationship between Jeju Island and Japan rather than that between Jeju and Gangwon-do Province. The divergence time between the maternal ancestor lineages of Japanese and Chinese populations was estimated to be 0.789±0.063 MYBP. The secondary divergence between Jeju and Japanese bats was calculated about to be 0.168±0.013 MYBP. The Jeju population has immigrated to the island at least fifty thousand years ago. In addition, ND1 haplotype analysis suggested that the insular bats have experienced at least two further genetic differentiation events within this island. Consequently, these findings suggested that the results of this study may play a critical role in understanding the phylogenetic relationship among East Asian bat populations of M. macrodactylus. To prepare more explainable information on evolutionary correlation, analysis is still required to examine using expanded samples from China, Russia, and southern parts of the Korean Peninsula.

Phylogenetic and Morphological Comparison between Thamnaconus septentrionalis and T. modestus Collected in Southwest Seashore (서남해에서 채집된 말쥐치 (Thamnaconus modestus)와 유사종 (T. septentrionalis)의 형태 및 계통유전학적 비교)

  • Yu, Tae-Sik;Park, Kiyun;Han, KyeongHo;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.229-239
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    • 2021
  • Thamnaconus modestus, distributed in the Northwest Pacific, has high economic value and is used in various seafood. In this study, the morphological and genetic characteristics of T. modestus and T. septentrionalis were compared and analyzed. We observed the external and internal morphology of T. modestus, sketched skeletal elements, and analyzed phylogenetic evolutionary relationships using the cytochrome c oxidase subunit I (COI) gene on mitochondrial DNA compared to T. septentrionalis. The T. modestus observed in this study had blackish-brown patterns irregularly scattered on the gray-brown body, and the fins were blue-green. Genetic analysis results based on the COI sequences of T. modestus showed seven types of base sequence variation; however, the homology was more than 98.8%. In addition, as a result of comparison of the COI nucleotide sequences and phylogenetic analysis in Tetraodontiformes, two T. septentrionalis sequences (JN813099, MW485059) were similar to T. modestus with 99% homology, and the other two T. septentrionalis sequences (EF607583, KP267619) were similar to those of species belonging to another genus Thamnaconus with 95% homology with T. modestus. It was not easy to classify the species based on morphological characteristics, and phylogenetic analysis between T. modestus and T. septentrionalis confirmed the difference in classification. These results provide the external and internal morphology of T. modestus and will be used as important information for the taxonomic study of T. modestus and T. septentrionalis.