• Title/Summary/Keyword: cytochrome oxidase subunit I

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Site-directed Mutagenesis of Five Conserved Residues of Subunit I of the Cytochrome cbb3 Oxidase in Rhodobacter capsulatus

  • Ozturk, Mehmet;Gurel, Ekrem;Watmough, Nicholas J.;Mandaci, Sevnur
    • BMB Reports
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    • v.40 no.5
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    • pp.697-707
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    • 2007
  • Cytochrome $cbb_3$ oxidase is a member of the heme-copper oxidase superfamily that catalyses the reduction of molecular oxygen to the water and conserves the liberated energy in the form of a proton gradient. Comparison of the amino acid sequences of subunit I from different classes of heme-copper oxidases showed that transmembrane helix VIII and the loop between transmembrane helices IX and X contain five highly conserved polar residues; Ser333, Ser340, Thr350, Asn390 and Thr394. To determine the relationship between these conserved amino acids and the activity and assembly of the $cbb_3$ oxidase in Rhodobacter capsulatus, each of these five conserved amino acids was substituted for alanine by site-directed mutagenesis. The effects of these mutations on catalytic activity were determined using a NADI plate assay and by measurements of the rate of oxygen consumption. The consequence of these mutations for the structural integrity of the $cbb_3$ oxidase was determined by SDS-PAGE analysis of chromatophore membranes followed by TMBZ staining. The results indicate that the Asn390Ala mutation led to a complete loss of enzyme activity and that the Ser333Ala mutation decreased the activity significantly. The remaining mutants cause a partial loss of catalytic activity. All of the mutant enzymes, except Asn390Ala, were apparently correctly assembled and stable in the membrane of the R. capsulatus.

DNA Barcoding of the Endangered Species Ellobium chinense (Mollusca, Gastropoda, Ellobiidae) from Coastal Areas of South Korea

  • Yi, Chang Ho;Jung, Tae Won;Kim, Il-Hun;Cho, In-Young;Kim, Min-Seop;Yoon, Moongeun;Kim, Won
    • Animal Systematics, Evolution and Diversity
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    • v.35 no.3
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    • pp.136-139
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    • 2019
  • The pulmonate gastropod Ellobium chinense (Pfeiffer, 1864) is an endangered marine species along the South Korean coasts due to habitat destruction and population declines. We sequenced the cytochrome c oxidase subunit 1 (COI) of 25 E. chinense specimens collected from five coastal sites in South Korea, and identified 16 unique haplotypes. The maximum intraspecific variation among individuals was 1.6%, while interspecific differences from another ellobiid species, Auriculastra duplicata (Pfeiffer, 1854), ranged from 21.9 to 23.0%. Our barcoding data will be useful to elucidate the phylogenetic relationships among pulmonate gastropods and infer the population genetic structure of E. chinense.

Genetic Diversity and Gene Flow Patterns in Pollicipes mitella in Korea Inferred from Mitochondrial DNA Sequence Analysis

  • Yoon, Moongeun;Jung, Ju-Yeon;Kim, Dong Soo
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.243-251
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    • 2013
  • Genetic diversity and gene flow patterns in Pollicipes mitella were investigated with a nucleotide sequence analysis of 514 base pairs from the mitochondrial cytochrome c oxidase subunit I gene (COI) in 124 samples collected from six Korean populations. In total, 59 haplotypes were defined by 40 variable nucleotide sites in the COI region. The haplotypes had shallow haplotype genealogy and no geographic associations. All populations had high haplotype diversity (0.909 to 0.979) and low nucleotide diversity (0.0055 to 0.0098). The haplotypes with recently diverged nucleotides were distributed by long-range larvae dispersal among regional populations. The pairwise fixation indices ($F_{ST}$) estimated with the exact test and migration rates indicate that substantial gene flow has occurred among populations as a result of sea currents, except between the Uljin (East Sea coast) and other Korean populations. This suggests that significant genetic differentiation and low migration rates have affected the Uljin population.

Amplified mitochondiral DNA identify four species of Tetranychus mites (Acarina: Tetranychidae) in Korea (미토콘드리아 DNA증폭을 이용한 한국의 잎응애속(Tetranychus;Acarina: Tetranychidae) 4종의 동정방법)

  • 이명렬;이문홍
    • Korean journal of applied entomology
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    • v.36 no.1
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    • pp.30-36
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    • 1997
  • Except for a cosmopolitan and major pest of apples, Tetranychus urticae Koch, Tetranychus mites in Korea such as T. viennensis Zaher, T. kanzawai Kishida, and T. truncams Ehara have been considered as quarantine pests by Canada and United States. Even though these mites are not feeders on apples, they are suspected to attach accidentally on apple h i t s in autumn as females enter the diapause. The characters used to identify Tetranychus mites have been confined to the shape of aedeagus in adult male. To develope a fast and accurate alternative identification protocol applied to hibernating female mites on apples, their mitochondrial DNA (mtDNA) were examined to find out any polymorphisms to discriminate each species from the other ones. Three pairs of primers for polymerase chain reaction (PCR) were used to amplify cytochrome oxidase subunit I (CO-I) coding region in mitochondrial DNA5 of four species of Tetranycus mites. The longest amplified product was estimated its size as about 680 bp. Digestion with restriction enzymes, AluI, Ddel, and Sau3A, showed length polymorphisms, which will he useful as diagnostic markers to identify Tetranychus mites. Schematic restriction maps in amplified region were shown for each species.

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DNA Barcoding of the Marine Protected Species Parasesarma bidens (Decapoda: Sesarmidea) from the Korean Waters

  • Kim, So Yeon;Yi, Chang Ho;Kim, Ji Min;Choi, Woo Yong;Kim, Hyoung Seop;Kim, Min-Seop
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.2
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    • pp.159-163
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    • 2020
  • Parasesarma bidens(De Haan, 1835) has been designated as a marine protected species by the Act on conservation and management of marine ecosystems. This crab has been recorded only from Jeju-do and Geomun-do, Republic of Korea. In this study, we describe for the first time the mitochondrial cytochrome c oxidase subunit I(COI) sequences of P. bidens. The intra-specific genetic distance among the Korean populations and between the Korean and Chinese populations ranged from 0% to 0.9% and 1.9% to 2.7%, respectively. The inter-specific genetic distances among the four Parasesarma species ranged from 10.9% to 12.8%. The finding of this study will be helpful to better describe P. bidens using COI DNA barcodes and can be used as basic data for their restoration and conservation research.

Estimation of micro-biota in the Upo wetland using eukaryotic barcode molecular markers

  • Park, Hyun-Chul;Bae, Chang-Hwan;Jun, Ju-Min;Kwak, Myoung-Hai
    • Journal of Ecology and Environment
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    • v.34 no.3
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    • pp.323-331
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    • 2011
  • Biodiversity and the community composition of micro-eukaryotic organisms were investigated in the Upo wetland in Korea using molecular analysis. Molecular identification was performed using cytochrome oxidase I (COI) and small subunit ribosomal DNA (SSU rDNA). The genomic DNA was isolated directly from soil samples. The COI and SSU rDNA regions were amplified using universal primers and then sequenced after cloning. In a similarity search of the obtained sequences with BLAST in the Genbank database, the closely related sequences from NCBI were used to identify the amplified sequences. A total of six eukaryotic groups (Annelida, Arthropoda, Rotifera, Chlorophyta, Bacillariophyta, and Stramenopiles) with COI and six groups (Annelida, Arthropoda, Rotifera, Alveolata, Fungi, and Apicomplexa) with SSU rDNA genes were determined in the Upo wetland. Among 38 taxa in 20 genera, which are closely related to the amplified sequences, 10 genera (50%) were newly reported in Korea and five genera (25%) were shown to be distributed in the Upo wetland. This approach is applicable to the development of an efficient method for monitoring biodiversity without traditional taxonomic processes and is expected to produce more accurate results in depositing molecular barcode data in the near future.

Sequence comparisons of 28S ribosomal DNA and mitochondrial cytochrome c oxidase subunit I of Metagonimus yokogawai, M. takahashii and M. miyatai

  • Lee, Soo-Ung;Huh, Sun;Sohn, Woon-Mok;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.42 no.3
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    • pp.129-135
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    • 2004
  • We compared the DNA sequences of the genus Metagonimus: M. yokogawai, M. takahashii, and M. miyatai. We obtained 288 D1 ribosomal DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (mtCOI) fragments from the adult worms by PCR, that were cloned and sequenced. Phylogenetic relationships inferred from the nucleotide sequences of the 28S D1 rDNA and mtCOI gene. M. takahashii and M. yokogawai are placed in the same clade supported by DNA sequence and phylogenie tree analysis in 28S D1 rDNA and mtCOI gene region. The above findings tell us that M. takahashii is closer to M. yokogawai than to M. miyatai genetically. This phylogenetic data also support the nomination of M. miyatai as a separate species.

DNA Barcoding of the Marine Proteced Species Pseudohelice subquadrata (Decapoda, Varunidae, Pseudohelice) from the Korean Waters

  • Kim, Ji Min;Kim, Jong-Gwan;Kim, So Yeon;Choi, Woo Yong;Kim, Hyung Seop;Kim, Min-Seop
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.3
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    • pp.228-231
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    • 2020
  • Pseudohelice subquadrata (Dana, 1851) is endangered due to its restricted habitat; hence, it has been designated as a marine protected species and endangered species by law in Korea. It has been recorded only Jeju-do and Geomun-do, Republic of Korea. The present study, is the first report on a cytochrome c oxidase subunit I DNA barcode for P. subquadrata. The maximum intra-specific genetic distance among all P. subquadrata individuals was found to be 0.5%, whereas inter-genetic distance within the same genus was 17.2-21.5% compared with Helice tientsinensis (Rathbun, 1931), H. tridens (De Haan, 1835), H. epicure (Ng et al., 2018), and Helicana wuana (Rathbun, 1931). Our barcoding data can thus be used as reference for restoration and conservation studies on P. subquadrata, which are designated as marine protected species.

Genetic diversity of spotted scat (Scatophagus argus) in Vietnam based on COI genes

  • Huy Van Nguyen;Minh Tu Nguyen;Nghia Duc Vo;Nguyen Thi Thao Phan;Quang Tan Hoang
    • Fisheries and Aquatic Sciences
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    • v.25 no.12
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    • pp.637-647
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    • 2022
  • A spotted scat, Scatophagus argus, has a high nutritional value and is among Asia's most widely consumed fish species. Thua Thien Hue's consumption market considers this species to be of high economic value and requires protection and conservation of the population. However, the studies on the identification and genetic diversity of S. argus distributed in Vietnam are still lacking. Therefore, mitochondrial cytochrome c oxidase subunit I (COI) gene was utilized to distinguish different populations and investigate the genetic diversity of two populations of S. argus from Tam Giang lagoon, Thua Thien Hue province (n = 31) and Ca Mau province (n = 14). The sequencing results indicated 13 distinct haplotypes among 45 sequences. Five single nucleotide polymorphisms were observed to distinguish Hue spotted scat population. The S. argus population in Ca Mau province was higher haplotype diversity (Hd) and nucleotide diversity (π) than those of Thua Thien Hue province, which demonstrates that there are minor differences between haplotypes. There were genetic distances ranging from 0%-4% within the populations and 6.67% between the two populations. In addition to the sequencing, the comparison of morphology, biology, culture, and the growth rate was sufficient to distinguish the spotted scat S. argus in Thua Thien Hue from Ca Mau.

Identification of eleven species of the Pleuronectidae family using DNA-based techniques

  • Eun-Mi Kim;Mi Nan Lee;Chun-Mae Dong;Eun Soo Noh;Young-Ok Kim
    • Fisheries and Aquatic Sciences
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    • v.26 no.11
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    • pp.678-688
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    • 2023
  • Flatfish are one of the largest families in the order Pleuronectiformes and are economically important edible marine fish species. However, they have similar morphological characteristics leading to challenges in classifying correctly, which may result in mislabeling and illegal sales, such as fraudulent labeling of processed food. Therefore, accurate identification is important to ensure the quality and safety of domestic markets in Korea. Species-specific primers were prepared from the mainly consumed eleven species of the order Pleuronectiformes. To rapidly identify the 11 flatfish species, a highly efficient, rapid, multiplex polymerase chain reaction (PCR) with species-specific primers was developed. Species-specific primer sets were designed for the mitochondrial DNA cytochrome c oxidase subunit I gene. Species-specific multiplex PCR (MSS-PCR) either specifically amplified a PCR product of a unique size or failed. This MSS-PCR analysis is easy to perform and yields reliable results in less time than the previous Sanger sequencing methods. This technique could be a powerful tool for the identification of the 11 species b the family Pleuronectidae and can contribute to the prevention of falsified labeling and protection of consumer rights.