• Title/Summary/Keyword: Sequence variation

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Identification of Entomopathogenic Fungus, Beauveria spp. F-101 Isolated from Thecodiplosis japonensis Using Internal Transcribed Spacer Sequence

  • Shin, Sang-Chul;Roh, Jong-Yul;Kim, Chul-Su;Park, Il-Kwon;Jeon, Mun-Jang;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.77-80
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    • 2004
  • For the development of the alternative control system against the major forest pests, Beauveria spp. F-101, isolated from a dead larva of Thecodiplosis japonensis, was selected because this isolate showed high pathogenicities against T. japonensis and Acantholyda parki. Beauveria spp. F-101 had irregular clustered conidio-phores and conidia borne on a distinctive apical zigzag extension, and it showed typical characteristic of the genus, Beauveria in morphology. For molecular based-identification, the ribosomal ITS region of Beauveria spp. F-101 was amplified with ITS1 and ITS4 primers, and cloned into pGEM- T Easy vector. The amplified PCR product was 569 bp in size and completely sequenced. The similarities of the cloned ITS sequence were 99 % and 97% to those of B. bassiana and B. brongniartii, respectively. In comparison to other species among the genus Beauveria, the ITS region of Beauveria spp. F-101 showed a similarity of 95% to B. amorpha, 95% to B. tenella, 89% to B. vermiconia and 69% to B. alba, respectively. In addition, in comparison to different genus, it had 95% similarities to Cordyceps militaris and 91% to Paecilomyces tenuipes. Accordingly, the current result suggests that Beauveria spp. F-101 was a variant of B. bassiana and it seems to be a new isolate considering sequence variation in ITS region.

Population Genetic Structure of the Bumblebee, Bombus ignitus (Hymenoptera: Apidae), Based on Mitochondrial COI Gene and Nuclear Ribosomal ITS2 Sequences

  • Oh, Hyung Keun;Yoon, Hyung Joo;Lee, Joo Young;Park, Jeong Sun;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.27 no.1
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    • pp.142-158
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    • 2013
  • The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is widely utilized for greenhouse pollination in South Korea. Understanding the magnitude of genetic diversity and geographic relationships is of fundamental importance for long term preservation and utilization. As a first step, we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to the "DNA barcode" region and the complete internal transcribed spacer 2 (ITS2) of nuclear ribosomal DNA from 88 individuals collected in nine South Korean localities. The complete ITS2 sequences were longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated 112-bp long repeats. The 658-bp long mtDNA sequences provided only six haplotypes with a maximum sequence divergence of 0.61% (4 bp), whereas the ITS sequences provided 84 sequence types with a maximum sequence divergence of 1.02% (21 sites). The combination of the current COI data with those of published data suggest that the B. ignitus in South Korea and China are genetically a large group, but those in Japan can be roughly separated into another group. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population were found to exist among the South Korean populations of B. ignitus, which suggests panmixia. This finding is consistent with our understanding of the dispersal capability of the species.

Intronic Sequences for the Discrimination of Silkworm Strains in Silkworm Powder

  • Kim, Kee-Young;Lee, Eun-Mee;Hong, Mee-Yeon;Kang, Pil-Don;Ryu, Kang-Sun;Kim, Ik-Soo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.2
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    • pp.93-100
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    • 2007
  • After a serial study on the therapeutic efficacy of the powdery silkworm for diabetics was positively resulted in, many powdery silkworm products were on the market in Korea. Up to now, however, no causal method is available to discriminate the strain of silkworms that is a major ingredient for manufacturing powdery silkworm, even though the quality of the powdery silkworm differs greatly by source and origin of strains. We previously were successful in identifying 25 silkworm strains kept in Korea using nine intronic sequences. In this study, we tested the utility of the nine intronic sequences to distinguish the most widely reared silkworm strains originated from Korea and China. Two intron regions, PTTH Intron3 and PTTH Intron3, showed a substantial sequence divergence (mean sequence divergence of 3.13% in PTTH Intron3 and 4.99% in PTTH Intron3). These two intronic sequences provided no identical sequences among the seven strains tested. Thus, these sequences each along can be used to discriminate the seven strains tested in this study. Furthermore, other intron regions, except for VDP Intron4 allowed us to discriminate $2{\sim}4$ strains by strain-specific unique insertion/deletion or substitution.

Genome-wide analysis of sequence variations in eight inbred watermelon lines (수박계통간 염색체수준의 유전적변이 분석)

  • Kim, Youn-Sung;Ko, Chan-Sup;Yang, Hee-Beom;Kang, Sun-Chul
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.164-173
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    • 2016
  • To investigate the genetic basis of phenotypic differences, sequence variations were analyzed in 8 inbred watermelon lines by re-sequencing. The number of sequence variations differed depending on the chromosome. Only 12.9% of SNPs were found within genes, whereas the rest were detected in promoter or intergenic regions. SNP density analysis showed that there was a highly variable region at the end of chromosome 6, which is similar to previously published findings. However, this region with high SNP density did not show much variation between the lines. In contrast, highly conserved regions with a size of 6.5-10 Mb were found in chromosomes 10 and 11. Pathway analysis suggested that the DIMBOA (a natural antibiotic)-glucoside degradation pathway was significantly different between the lines, indicating that the eight lines may have different levels of pathogen resistance. Among the carbohydrate-related genes, the alpha-galactosidase gene was the most variable among the lines. Information from this study will be helpful in understanding the watermelon breeding process at the molecular level.

Substrate Specificity of the Yeast Protein Tyrosine Phosphatase, PTP1, Overexpressed from an Escherichia coli Expression System

  • Kwon, Mi-Yun;Oh, Min-Su;Han, Jun-Pil;Cho, Hyeong-Jin
    • BMB Reports
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    • v.29 no.4
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    • pp.386-392
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    • 1996
  • A Saccharomyces cerevisiae Protein Tyrosine Phosphatase, PTP1, was expressed from an Escherichia coli expression system and milligram quantities of active PTP1 were purified chromatographically. The substrate specificity of the recombinant PTP1 was probed using synthetic phosphotyrosine-containing peptides corresponding to the regulatory phosphorylation sites of the yeast MAP kinase homologues $Fus3_{176-186}$, $Kss1_{179-189}$, and $Hog1_{170-180}$. Peptide sequences derived from the MAP kinase homologues were chosen arbitrarily as starting points for sequence variation studies even though they are not likely to be candidates for physiological substrates of PTP1. Phosphotyrosyl-$Hog1_{170-180}$ peptide showed a $K_M$ value of 877 ${\mu}M$ and phosphorylated $Kss1_{179-189}$ and $Fus3_{176-186}$ peptides showed lower $K_M$ values of 74 ${\mu}M$ and 51 ${\mu}M$ each. To study the effect of sequence variations of the peptide, amino acids of the undecapeptide $Hog1_{170-180}$ (DPQMTGpYVSTR) were sequentially substituted by an alanine residue. More extensive variations of each amino acid revealed positional importance of each amino acid residue. Based on these results, we derived a peptide sequence (DADEpYDA) that is recognized by PTP1 with an affinity ($K_M$ is 4 ${\mu}M$) significantly higher than that of the peptides derived from the phosphorylation sites of Fus3, Kss1, and Hog1.

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CNVR Detection Reflecting the Properties of the Reference Sequence in HLA Region (레퍼런스 시퀀스의 특성을 고려한 HLA 영역에서의 CNVR 탐지)

  • Lee, Jong-Keun;Hong, Dong-Wan;Yoon, Jee-Hee
    • Journal of KIISE:Computing Practices and Letters
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    • v.16 no.6
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    • pp.712-716
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    • 2010
  • In this paper, we propose a novel shape-based approach to detect CNV regions (CNVR) by analyzing the coverage graph obtained by aligning the giga-sequencing data onto the human reference sequence. The proposed algorithm proceeds in two steps: a filtering step and a post-processing step. In the filtering step, it takes several shape parameters as input and extracts candidate CNVRs having various depth and width. In the post-processing step, it revises the candidate regions to make up for errors potentially included in the reference sequence and giga-sequencing data, and filters out regions with high ratio of GC-contents, and returns the final result set from those candidate CNVRs. To verify the superiority of our approach, we performed extensive experiments using giga-sequencing data publicly opened by "1000 genome project" and verified the accuracy by comparing our results with those registered in DGV database. The result revealed that our approach successfully finds the CNVR having various shapes (gains or losses) in HLA (Human Leukocyte Antigen) region.

A Study on the 3D Shape Reconstruction Algorithm of an Indoor Environment Using Active Stereo Vision (능동 스테레오 비젼을 이용한 실내환경의 3차원 형상 재구성 알고리즘)

  • Byun, Ki-Won;Joo, Jae-Heum;Nam, Ki-Gon
    • Journal of the Institute of Convergence Signal Processing
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    • v.10 no.1
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    • pp.13-22
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    • 2009
  • In this paper, we propose the 3D shape reconstruction method that combine the mosaic method and the active stereo matching using the laser beam. The active stereo matching method detects the position information of the irradiated laser beam on object by analyzing the color and brightness variation of left and right image, and acquires the depth information in epipolar line. The mosaic method extracts feature point of image by using harris comer detection and matches the same keypoint between the sequence of images using the keypoint descriptor index method and infers correlation between the sequence of images. The depth information of the sequence image was calculated by the active stereo matching and the mosaic method. The merged depth information was reconstructed to the 3D shape information by wrapping and blending with image color and texture. The proposed reconstruction method could acquire strong the 3D distance information, and overcome constraint of place and distance etc, by using laser slit beam and stereo camera.

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Morphological Variation and Partial Mitochondrial Sequence Analysis of Echinoid Species from the Coasts of the East Sea (동해 연안에 서식하는 성게의 형태변이와 미토콘드리아 유전자 분석)

  • Shin, Ji-Hye;Kim, Sung-Gyu;Kim, Young-Dae;Sohn, Young-Chang
    • Journal of Aquaculture
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    • v.21 no.3
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    • pp.139-145
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    • 2008
  • Morphological classification of echinoid species has many difficulties because of their phenotypic variations. In the present study, we analyzed morphotypes and partial mitochondrial 12S rDNA sequences of four sea urchin species classified as Pseudocentrotus depressus, Anthocidaris crassispina, Hemicentrotus pulcherrimus and Strongylocentrotus nudus, and unidentified four species collected from the coasts of the East sea. Their genomic DNAs were extracted from gonads and mitochondrial 12S rDNA sequences were amplified by the polymerase chain reaction (PCR) method. The sequence identities among the known four sea urchin species were 87.4-95.6%. The sequence identities among the unidentified four species were 99.4-99.6% and showed the highest homology to S. intermedius(99.8%). Thus, our phylogenetic tree indicates that the unidentified four species belong to S. intermedius.

Phylogenetic Relationship in Different Commercial Strains of Pleurotus nebrodensis Based on ITS Sequence and RAPD

  • Alam, Nuhu;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyeong-Gyun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • v.37 no.3
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    • pp.183-188
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    • 2009
  • The molecular phylogeny in nine different commercial cultivated strains of Pleurotus nebrodensis was studied based on their internal transcribed spacer (ITS) region and RAPD. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 592 to 614 bp. The size of ITS1 and ITS2 regions varied among the strains from 219 to 228 bp and 211 to 229 bp, respectively. The sequence of ITS2 was more variable than ITS1 and the region of 5.8S sequences were identical. Phylogenetic tree of the ITS region sequences indicated that selected strains were classified into five clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by RAPD with 20 arbitrary primers. Twelve primers were efficient to applying amplification of the genomic DNA. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD and ITS analysis techniques were able to detect genetic variation among the tested strains. Experimental results suggested that IUM-1381, IUM-3914, IUM-1495 and AY-581431 strains were genetically very similar. Therefore, all IUM and NCBI gene bank strains of P. nebrodensis were genetically same with some variations.

Genetic Variation of Scytalone Dehydratase Gene in Colletotrichum spp. (Colletotrichum spp. 유래 scytalone dehydratase 유전자의 유전적 다양성 비교)

  • Yun, Yeo-Hong;Hyeon, Min-U;Suh, Dong-Yeon;Kim, Yong-Min;Kim, Seong-Hwan;Choi, Chang-Won;Kim, Young-Shick
    • The Korean Journal of Mycology
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    • v.37 no.2
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    • pp.155-160
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    • 2009
  • The production of dihydroxynaphthalene (DHN) melanin is known to be essential factor for pathogenicity in Colletotrichum lagenarium. However, the genetic diversity of melanin genes was not much known among Colletotrichum spp. To investigate the variability of melanin gene in Colletotrichum spp. that cause anthracnose on diverse crops including tomato, we cloned and sequenced partial sd, one of DHN melanin genes encoding for scytalone dehydratase, from eight strains of C. coccodes, C. acutatum, C. truncatum C. caricae, and C. musae. The size of PCR-amplified sd ranged 437 bp to 545 bp. The nucleotide sequence identity of sd among the Colletotrichum strains tested varied from 49% to 99%. All of the PCR-amplified sd from eight strains contain an intron and have two exons coding for 122 amino acids. Overall, the size and nucleotide sequence of sd varied among the five Colletotrichum spp. Sequence identity of the predicted scytalone dehydratase protein of 122 amino acids ranged 50 to 99%. Phylogentic analysis based on the sd nucleotide sequences revealed that the five Colletotrichum spp. could be genetically divided.