• Title/Summary/Keyword: COI 유전자

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Mitochondrial DNA Swquence Variation of the Firefly, Pyrocoelia rufa(Coleoptera: Lampyridae), in Korea (늦반딧불이 Pyrocoelis rufa(딱정벌레목: 반딧불이과)의 미토콘드리아 DNA 염기서열 변이)

  • 이상철;김익수;배진식;진병래;김삼은;김종길;윤형주;양성렬;임수호
    • Korean journal of applied entomology
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    • v.39 no.3
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    • pp.181-191
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    • 2000
  • We have sequenced a portion of mitochondrial CO! gene (403 bp) of the firefly, Pyrocoelia rufa, to investigate genetic diversity within population, geographic variation, and phylogenetic relationships among haplotypes. A total of seven mtDNA haplotypes ranging in sequence divergence from 0.2% to 1.2% were obtained from 26 fireflies collected at four localities in Korea: Namhae, Pusan, Muju, and Yongin. The samples collected at the urban area, Pusan, were all fixed with one haplotype, differently those collected at the forest and/or agricultural areas. This appears to suggest that habitat fragmentation and population bottleneck caused by urbanization might have been severe in Pusan. On the other hand, from Muju known as the largest habitat and sanctuary for the firefly, four haplotypes with the maximum sequence divergence of 1.0% were obtained, and this estimate was the highest among the areas studied. The fireflies collected at the isolated islet, Namhae, revealed relatively low haplotype diversity(H=0.25), but one haplotype (PR7) was phylogenetically differentiated from others. This phenomenon was explained in terms of biogeographic history of the island and gene flow in the recent past. Grouping of Muju- Y ongin and Pusan-Namhae, respectively, in the hierarchical genetic analysis suggests the presence of historically occurred, biogeographic barrier against gene flow between them.

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The Population Genetic Structure of the Oyster Crassostera gigas (Bivalvia:Ostreidae) from Gamak Bay in Korea (가막산 참굴의 집단 구조 분석)

  • Cho, Eun-Seob;Jeong, Hee-Dong
    • Journal of Life Science
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    • v.18 no.7
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    • pp.1015-1018
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    • 2008
  • To analyze the population genetic structure of the oyster Crassostrea gigas Thunberg, 34 specimens werecollected from Gamak bay in March, 2007. Total genomic DNA was extracted from each sample and PCR was performed to identify haplotypes of oyster by using HCO2918 and LCO1491 primers. Four kinds of haplotypes (CR1, CR2, CR3, and CR4) were identified. Among these group, CR3 showed the highest relative frequency at 73% than any other of haplotypes. On the basis of hierarchical genetic structure, the population of Gamak showed a higher genetic relationship with Namhae, but the genetic distance between southern and western coasts was negative and no statistical significance was found (p>0.05). Consequently, the oyster from Korea coast is determined to be both homogenous and large.

Identification of the South Korean Hermit Beetle (Coleoptera: Scarabaeidae: Cetoniinae) (남한산 큰자색호랑꽃무지의 종 동정 (딱정벌레목, 풍뎅이과, 꽃무지아과))

  • Han, Taeman;Park, In Gyun;Kim, Ki-Kyoung;Ivanov, Sergey;Park, Haechul
    • Korean journal of applied entomology
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    • v.56 no.3
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    • pp.229-239
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    • 2017
  • The South Korean hermit beetle has previously been identified as Osmoderma opicum, which is distributed in Japan. Because of its rarity, this species is classified as an endangered species. To date, however, the identity of this species in South Korea has not been conclusively confirmed. To assess the taxonomic status of the hermit beetle occurring in South Korea, we performed a comparative study with the beetle's Eurasian congeners, based on morphological examination and molecular analysis using COI gene sequences. The results clearly showed that the South Korean hermit beetle is identical to Osmoderma caeleste, which has been described from the Russian Far East. Therefore, we suggest that the taxonomic identity of the South Korean hermit beetle is O. caeleste.

Development of Ultra-rapid Nested PCR Method for Detection of Specific Gene of Tracheal Mite (Acarapis woodi) (기문응애(Acarapis woodi) 특이 유전자 검출을 위한 초고속 nested PCR법 개발)

  • Kim, MoonJung;Kim, Byoung-Hee;Kim, SoMin;Truong, A Tai;Kim, Jung-Min;Kim, Seonmi;Yoon, Byoung-Su
    • Journal of Apiculture
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    • v.34 no.1
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    • pp.15-26
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    • 2019
  • Tracheal mite (Acarapis woodi) is an internal parasite that is parasitic on the bronchus of adult bees and sucks fluid from the trachea. Since its first report by Rennie, it has been spread throughout Europe and in some Asian regions, with adjacent Japan and China reported in 2011 and 2012, respectively. Korea detected specific genes of A. woodi in 2015, but only one of 99 samples has been identified and the being of A. woodi has not been confirmed. In this study, we established a specific nested PCR method to confirm for detecting low-copy number of A. woodi-specific gene in bee samples. As a result, A. woodi-specific COI gene was amplified in 15 of 23 samples, and they were judged positive by melting point analysis and sequencing analysis. Although we could not observe the existence of the mites in bees, our results suggest that tracheal mit might exist in nature.

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.

Population Genetic Structure of the Korean Endemic Species, Iksookimia pacifica (Pisces: Cobitidae) Distributed in Northeast Korea (한국고유종 북방종개(어류강, 미꾸리과)의 집단유전학적 구조)

  • Jang, Sook-Jin;Ko, Myeong-Hun;Kwan, Ye-seul;Won, Yong-Jin
    • Korean Journal of Environment and Ecology
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    • v.31 no.5
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    • pp.461-471
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    • 2017
  • Population genetic studies of 10 groups of Iksookimia pacifica were conducted to investigate the genetic diversity and population genetic structure across its known range in South Korea. Population DNA sequences of one mitochondrial gene (mtCOI) and three nuclear genes (IRBP, EGR2B, RAG1) were examined in samples collected from ten streams that flow into the East Sea. Both mitochondrial and nuclear sequences exhibited significant differentiation among populations except a few cases. The Bayesian analysis of the multi-locus genotypes inferred from the DNA sequences of nuclear genes clustered the individual fish largely into two geographical groups: a northern group (from Baebong stream to Cheonjin stream) and a southern group (Yangyangnamdae stream to Gangneungnamdae stream). Given that the streams flowing into the East Sea are geographically isolated water systems, such separation of genotypes can be interpreted by the geographical separation of common ancestors into north and south that had colonized South Korea. Since the initial geographical separation of the ancestral population by north and south, the ancestral groups seem to have experienced further differentiation into the current genetic clusters through the physical isolation of streams by the East Sea in each region. It is notable that many individuals in the Jasan stream formed a genetic cluster with those of Yangyangnamdae and Gangneungnamdae streams which are distant from each other. In addition, mitochondrial gene showed low genetic differentiation between some neighboring populations and very low level of genetic diversity in several populations. The present population genetic study will provide valuable information for the conservation and management of the Korean endemic fish species, I. paicifica.

Application for Identification of Food Raw Materials by PCR using Universal Primer (일반 프라이머를 이용한 PCR의 식품원료 진위 판별에 적용)

  • Park, Yong-Chjun;Jin, Sang-Ook;Lim, Ji-Young;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae;Lee, Sang-Jae;Lee, Kwang-Ho;Yoon, Hae-Seong
    • Journal of Food Hygiene and Safety
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    • v.27 no.3
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    • pp.317-324
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    • 2012
  • In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

Development of Species-Specific PCR to Determine the Animal Raw Material (종 특이 프라이머를 이용한 동물성 식품원료의 진위 판별법 개발)

  • Kim, Kyu-Heon;Lee, Ho-Yeon;Kim, Yong-Sang;Kim, Mi-Ra;Jung, Yoo Kyung;Lee, Jae-Hwang;Chang, Hye-Sook;Park, Yong-Chjun;Kim, Sang Yub;Choi, Jang Duck;Jang, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.347-355
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    • 2014
  • In this study, the detection method was developed using molecular biological technique to distinguish authenticity of animal raw materials. The genes for distinction of species about animals targeted at Cytochrome c oxidase subunit I (COI), Cytochrome b (Cytb), and 16S ribosomal RNA (16S rRNA) genes in mitochondrial DNA. The species-specific primers were designed by that Polymerase Chain Reaction (PCR) product size was around 200 bp for applying to processed products. The target 24 raw materials were 2 species of domestic animals, 6 species of poultry, 2 species of freshwater fishes, 13 species of marine fishes and 1 species of crustaceans. The results of PCR for Rabbit, Fox, Pheasant, Domestic Pigeon, Rufous Turtle Dove, Quail, Tree Sparrow, Barn Swallow, Catfish, Mandarin Fish, Flying Fish, Mallotus villosus, Pacific Herring, Sand Lance, Japanese Anchovy, Small Yellow Croaker, Halibut, Jacopever, Skate Ray, Ray, File Fish, Sea Bass, Sea Urchin, and Lobster raw materials were confirmed 113 bp ~ 218 bp, respectively. Also, non-specific PCR products were not detected in compare species by species-specific primers. The method using primers developed in this study may be applied to distinguish an authenticity of food materials included animal raw materials for various processed products.

Thrips Infesting Hot Pepper Cultured in Greenhouses and Variation in Gene Sequences Encoded in TSWV (시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이)

  • Kim, Chulyoung;Choi, Duyeol;Kang, Jeong Hun;Ahmed, Shabbir;Kil, Eui-Joon;Kwon, Gimyeon;Lee, Gwan-Seok;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.60 no.4
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    • pp.387-401
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    • 2021
  • Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

Development of the Duplex PCR Method of Identifying Trachurus japonicus and Trachurus novaezelandiae (다중 PCR 분석법을 이용한 전갱이속 어종의 신속한 종판별 분석법 개발)

  • Park, Yeon Jung;Lee, Mi Nan;Kim, Eun Mi;Noh, Eun Soo;Noh, Jae Koo;Park, Jung Youn;Kang, Jung-Ha
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1062-1067
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    • 2018
  • Reliable labeling of fish products can reassure consumers regarding the identity and quality of seafoods. Therefore, techniques that can identify adulteration or mislabeling are valuable. To rapidly identify two Trachurus species, Trachurus japonicus and Trachurus novaezelandiae, a highly efficient, rapid, duplex polymerase chain reaction (PCR) having two species-specific primers simultaneously was identified. This species-specific primer focused on a single nucleotide mismatch in the 3'-terminal base of a primer designed in the mitochondrial cytochrome c oxidase (COI) subunit I DNA. To optimize the duplex PCR condition, gradient PCR reactions were conducted to determine the primer annealing temperature and the primer concentration. The PCR's product was observed on the gel, suggesting that DNA molecules may be useful in differentiating the two species. The length of the amplification fragments were 103 bp for Trachurus japonicus and 214 bp for Trachurus novaezelandiae, which, along with the species-specific primer visualized by agarose gel electrophoresis, enabled accurate distinction of the species of the Trachurus genus. These results indicate that the duplex PCR, which has a species-specific primer based on single nucleotide polymorphism (SNP), can be useful for rapidly differentiating the two species of Trachurus. This duplex PCR analysis is simple, rapid, and reliable, and could be beneficial to protecting consumers' rights.