• Title/Summary/Keyword: Gene Identification

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Cloning and Phylogenetic Analysis of Chitin Synthase Gene from Entomopathogenic Fungus, Beauveria brongniartii

  • Nam, Jin-Sik;Lee, Dong-Hun;Park, Ho-Yong;Bae, Kyung-Sook
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.222-227
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    • 1997
  • DNA fragments homologous to chitin synthase gene were amplified from the genomic DNA of Beauveria brongniartii by PCR using degenerate primers. Cloning and sequencing of the PCR-amplified fragments led to the identification of a gene, designated BbCHSl. Comparison of the deduced amino acid sequence of BbCHSl with those of other Euascomycetes revealed that BbCHSl is a gene for class II chitin synthase. The Blastp search of the deduced amino acid sequence of BbCHSl displayed the highest rate of similarity, 95.8%, with CHS2 of Metarhizium unisopliae. Phylogenetic analysis of the amino acid sequences confirmed the taxonomic and evolutionary position of B. brongniartii, which was previously derived by traditional fungal classification based on morphological features.

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Classification in Different Genera by Cytochrome Oxidase Subunit I Gene Using CNN-LSTM Hybrid Model

  • Meijing Li;Dongkeun Kim
    • Journal of information and communication convergence engineering
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    • v.21 no.2
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    • pp.159-166
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    • 2023
  • The COI gene is a sequence of approximately 650 bp at the 5' terminal of the mitochondrial Cytochrome c Oxidase subunit I (COI) gene. As an effective DeoxyriboNucleic Acid (DNA) barcode, it is widely used for the taxonomic identification and evolutionary analysis of species. We created a CNN-LSTM hybrid model by combining the gene features partially extracted by the Long Short-Term Memory ( LSTM ) network with the feature maps obtained by the CNN. Compared to K-Means Clustering, Support Vector Machines (SVM), and a single CNN classification model, after training 278 samples in a training set that included 15 genera from two orders, the CNN-LSTM hybrid model achieved 94% accuracy in the test set, which contained 118 samples. We augmented the training set samples and four genera into four orders, and the classification accuracy of the test set reached 100%. This study also proposes calculating the cosine similarity between the training and test sets to initially assess the reliability of the predicted results and discover new species.

Identification of Cambodian Gnetum (Gnetaceae, Gnetales) species by DNA barcoding

  • Kim, Joo Hwan;Won, Hyosig
    • Korean Journal of Plant Taxonomy
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    • v.46 no.2
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    • pp.163-174
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    • 2016
  • Gnetum (Gnetaceae, Gnetales) is a gymnosperm genus with ca. 35 species distributed in tropical forests around the world. Due to its dioecious habit and lack of diagnostic characters from vegetative tissue, the identification of Gnetum species is not easy without seeds or reproductive structures. To identify and verify their phylogenetic positions, we applied DNA barcoding to Cambodian Gnetum collections gathered between 2010 and 2015, with previously designed cp matK gene primers. We newly sequenced partial matK sequences from 72 Gnetum collections, 43 out of 72 from Cambodia, and analyzed 115 Gnetum accessions using the neighbor-joining method. The resulting neighbor-joining tree categorized Cambodian Gnetum samples into three clades of species: G. macrostachyum, G. montanum, and G. aff. gracilipes. The recognition of G. aff. gracilipes in Cambodia is reported here for the first time. Taxonomic information for the three recognized Cambodian Gnetum species is provided and the benefits of the taxonomic reevaluation assisted by DNA barcoding are emphasized in this work.

Rapid Identification of Potato Scab Causing Streptomyces spp. from Soil Using Pathogenicity Specific Primers

  • Kim, Jeom-Soon;Lee, Young-Gyu;Ryu, Kyoung-Yul;Kim, Jong-Tae;Cheon, Jeong-Uk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.134.2-135
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    • 2003
  • The plant-pathogenic species S. scabies, S. acidiscabies, and S. turgidiscabies cause the scab disease of potato and produce the phytotoxins, thaxtomins. necl, a gene conferring a necrogenic phenotype, is involved in pathogenicity and physically linked to the thaxtomin A biosynthetic genes. Identification of the pathogenic strains of Streptomyces from soil was performed through the polymerase chain reaction by using specific pathogenicity primer sets derived from the necl gene sequences of Streptomyces smbies. The DNA was extracted from soil using a bead-beating machine and modifications of the FastPrep system. The DNA was suitable for direct use in the PCR. The PCR products showed the bands of approximately 460 bp. This methods can be very usuful in identifying species responsible for scab diseases and studying on the ecology of plant-pathogenic Streptomyces spp.

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Identification and Characterization of Thermoplasma acidophilum 2-Keto-3-Deoxy-D-Gluconate Kinase: A New Class of Sugar Kinases

  • Jung, Jin-Hwa;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.535-539
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    • 2005
  • The thermoacidophilic archaeon Thermoplasma acidophilum has long been known to utilize D-glucose via the non-phosphorylated Entner-Doudoroff (nED) pathway. We now report the identification of a gene encoding 2-keto-3-deoxy-D-gluconate (KDG) kinase. The discovery of this gene implies the presence of a glycolysis pathway, other than the nED pathway. It was found that Ta0122 in the T. acidophilum genome corresponded to KDG kinase. This enzyme shares no similarity with known KDG kinases, and belongs to a novel class of sugar kinases. Of the five sugars tested only KDG was utilized as a substrate.

Identification of Four Cyst Nematodes using PCR-RFLP in Korea (PCR-RFLP를 이용한 국내 분포 씨스트선충 4종의 동정)

  • Ko, Hyoung-Rai;Kang, Heonil;Park, Eun-Hyoung;Kim, Eun-Hwa;Lee, Jae-Kook
    • Korean Journal of Organic Agriculture
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    • v.27 no.3
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    • pp.353-363
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    • 2019
  • To identify four cyst nematodes (Heterodera schachtii, H. trifolii, H. glycines, H. sojae) that are economically important plant-parasitic nematodes in Korea, restriction fragment length polymorphism (RFLP) by 8 endonucleases (PstI, VspI, AlwI, RsaI, MvaI, EcoRI, Eco72I, Hinf I) was performed based on sequence difference of mitochondrial DNA cytochrome c oxidase subunit I (COI) gene. As a result, species-specific DNA band patterns by RsaI endonuclease were observed in H. schachtii. The specific patterns was in H. trifolii by 3 endonucleases (VspI, AlwI, Hinf I), and was in H. glycines by Hinf I. While, H. sojae was not digested by 4 endonuclease (VspI, AlwI, RsaI, Hinf I). This study showed that four cyst nematodes could be distinguished using RFLP by 4 endonucleases (RsaI, VspI, AlwI, Hinf I) based on the sequence difference of COI gene.

Differentiation of three scuticociliatosis causing species in olive flounder (Paralichthys olivaceus) by multiplex PCR

  • Kim, Sung Mi;Lee, Eun Hye;Kim, Ki Hong
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.145-149
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    • 2004
  • The definitive identification of ciliate species by morphological characteristics relies on time-consuming and laborious staining techniques. Therefore, in this study, we discriminated 3 scuticociliatosis causing species - Pseudocohnilembus persalinus, Uronema marinum and Philasterides dicentrarchi - in cultured olive flounder by multiplex PCR. The multiplex PCR based on the species-specific amplification of small subunit ribosomal RNA (SS rRNA) gene sequence enabled us to distinguish the 3 scuticociliate species in a simple and rapid manner, even in the sample containing the three species simultaneously. These data suggest that the multiplex PCR strategy would make it possible to avoid the cumbersome and time-consuming procedures of morphological analysis for the definitive identification of scuticociliates.

Rapid Identification of Potato Scab Causing Streptomyces spp. Using Pathogenicity Specific Primers

  • Shin, Pyung-Gyun;Kim, Jeom-Soon;Hahm, Young-Il
    • The Plant Pathology Journal
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    • v.18 no.6
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    • pp.338-341
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    • 2002
  • The potato scab is caused by several species of Streptomyces. Among these species, only pathogenic strains were found to produce thaxtomin A characterized by necrotic bioassay and HPLC. In this study, identification of the pathogenic strains of Streptomyces was performed through the polymerase chain reaction (PCR) by using specific pathogenicity primer sets derived from the nec1 gene sequences of Streptomyces scabies. The expected PCR products were obtained approximately 580 bp and confirmed by sequencing. This PCR technique can be used effectively to identify the pathogenic Streptomyces species, that cause scab on potato tubers.

Molecular identification of Anisakid worm third stage larvae isolated from masou salmon Oncorhynchus masou (시마연어, Oncorhynchus masou에서 분리된 아니사키스 속 선충 3기 유충의 분자생물학적 방법을 이용한 동정)

  • Jeon, Chan-Hyeok;Setyobudi, Eko;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.23 no.3
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    • pp.421-427
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    • 2010
  • Anisakid nematodes third stage larvae were isolated from the muscles of masou salmon (Oncorhynchus masou). Fish were purchased from Jumunjin fishery market in Gangneung. Four Anisakid third stage larvae were isolated from 4 fish. Molecular identification of the isolated worms was conducted by PCR-RFLP analysis of ribosomal DNA internal transcribed spacer region and direct sequencing of mitochondrial DNA cox2 gene. As results, all the tested individual worms were identified as Anisakis simplex (sensu stricto). This is the first report of molecular detection of anisakid worms in salmonid fishes in Korea.

Multiplex TaqMan qPCR Assay for Detection, Identification, and Quantification of Three Sclerotinia Species

  • Dong Jae Lee;Jin A Lee;Dae-Han Chae;Hwi-Seo Jang;Young-Joon Choi;Dalsoo Kim
    • Mycobiology
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    • v.50 no.5
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    • pp.382-388
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    • 2022
  • White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.