• Title/Summary/Keyword: Molecular sequence analyses

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Morphology, Pathogenicity and Molecular analysis of Alternaria Isolates from Solanaceous Crops (oral)

  • Cho, H.S.;Park, M.S.;Kim, B.R.;Yu, S.H.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.112.3-113
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    • 2003
  • More than 30 isolates of Alternaria were obtained from various solanaceous crops in Korea. For all isolates, morphological characteristics of the conidia were determined and compared with those of representative isolates of A. solani and A. tomatophila. Pathogenicity test was performed to Potato, tomato, egg plant and red Pepper and molecular characteristics of them including the representative isolates were determined using sequence analyses of ITS rDNA and histone H3 gene, and URP-PCR analysis. Based on morphological characteristics, the isolates from the solanaceous crops were grouped as identical or very similar to either A. tomatophila(ATO), A. solani(ASO), and unidentified Altemaria sp.(ASP). Among the molecular markers used in this study, the URP-PCR analysis was found to be appropriate for taxonomic resolution of these species. Based on the conidial morphology, pathogenicity test and molecular characteristics, A. tomatophila(early blight of tomato) could be distinguished from A. solani(early blight of potato), and the Alternaria sp.(ASP) from potato, which was closely related to A. solani in conidial morphology, was considered as a new species.

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First Molecular Characterization of Hypoderma actaeon in Cattle and Red Deer (Cervus elaphus) in Portugal

  • Ahmed, Haroon;Sousa, Sergio Ramalho;Simsek, Sami;Anastacio, Sofia;Kilinc, Seyma Gunyakti
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.653-658
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    • 2017
  • Hypoderma spp. larvae cause subcutaneous myiasis in several animal species. The objective of the present investigation was to identify and characterize morphologically and molecularly the larvae of Hypoderma spp. collected from cattle (Bos taurus taurus) and red deer (Cervus elaphus) in the district of Castelo Branco, Portugal. For this purpose, a total of 8 larvae were collected from cattle (n=2) and red deer (n=6). After morphological identification of Hypoderma spp. larvae, molecular characterization was based on PCR-RFLP and mitochondrial CO1 gene sequence analysis. All larvae were morphologically characterized as the third instar larvae (L3) of H. actaeon. Two restriction enzymes were used for molecular identification of the larvae. TaqI restriction enzyme was not able to cut H. actaeon. However, MboII restriction enzyme differentiated Hypoderma species showing 210 and 450 bp bands in H. actaeon. Furthermore, according to the alignment of the mt-CO1 gene sequences of Hypoderma species and to PCR-RFLP findings, all the identified Hypoderma larvae were confirmed as H. actaeon. This is the first report of identification of Hypoderma spp. (Diptera; Oestridae) from cattle and red deer in Portugal, based on morphological and molecular analyses.

Molecular Phylogeny and Divergence Time Estimation of the Soft Coral Dendronephthya gigantea (Alcyonacea: Nephtheidae)

  • Kim, Boa;Kong, So-Ra;Song, Jun-Im;Won, Yong-Jin
    • Animal Systematics, Evolution and Diversity
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    • v.24 no.3
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    • pp.327-332
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    • 2008
  • Soft coral Dendronephthya gigantea (Verrill, 1864) is a conspicuous species dominating shallow sea waters of Jejudo Island, Korea. Recently its whole mitochondrial genome sequencing was completed by us and the sequence information provided an opportunity to test the age of Octocorallia and time of evolutionary separation between some representative orders of the subclass Octocorallia. Molecular phylogenetic analyses based on 13 mitochondrial protein encoding genes revealed a polyphyletic relationship among octocorallians representing two orders (Alcyonacea and Gorgonacea) and four families (Alcyoniidae, Nephtheidae, Briareidae, and Gorgoniidae). Estimates of divergence times among octocorallians indicate that the first splitting might occur around end of or after Cretaceous period (50-79 million years ago (Ma)). The age is relatively young compared to the long history of stony sea corals (>240 Ma). Taken together our result suggests a possible relatively recent radiating evolution at least in the order Alcyonacea and Gorgonacea. Molecular dating and phylogenetic analysis based on much broader taxon sampling and many genes might give an insight into this interesting hypothesis.

Unveiling the Diversity of Hydnum in the Republic of Korea with One New Species, Hydnum paucispinum

  • Ji Seon Kim;Wonjun Lee;Changmu Kim;Hanna Park;Chang Sun Kim;Young Woon Lim
    • Mycobiology
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    • v.51 no.5
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    • pp.300-312
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    • 2023
  • Hydnum is a genus of ectomycorrhizal fungi belonging to the Hydnaceae family. It is widely distributed across different regions of the world, including North America, Europe, and Asia; however, some of them showed disjunct distributions. In recent years, with the integration of molecular techniques, the taxonomy and classification of Hydnum have undergone several revisions and advancements. However, these changes have not yet been applied in the Republic of Korea. In this study, we conducted an integrated analysis combining the morphological and molecular analyses of 30 specimens collected over a period of approximately 10 years in the Republic of Korea. For molecular analysis, the sequence data of the internal transcribed spacer (ITS) region, the large subunit of nuclear ribosomal RNA gene (nrLSU), and a portion of translation elongation factor 1-a (TEF1) were employed as molecular markers. Through this study, we identified eight species that had previously not been reported to occur in the Republic of Korea, including one new species, Hydnum paucispinum. A taxonomic key and detailed descriptions of the eight Hydnum species are provided in this study.

Nucleotide Sequence Analyses of p10 Gene and its Promoter of Hyphantria cunea Nuclear Polyhedrosis Virus (Hyphantria cunea Nuclear Polyhedrosis Virus p10유전자와 프로모터의 염기서열 결정)

  • Park, Sun-A;Cha, Sung-Chul;Chang, Jae-Hyeok;Lee, Hyung-Hoan
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.131-137
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    • 1996
  • The sequences of p10 gene its promoter of Hyphantria cunea NPV were determined. According to the sequence analysis, the putative p10 gene ORF has 285 bp. The 5'-non-coding leader sequence of the p10 gene promoter contained the TATA box and the putative transcription initiation site TAAG motif. Poly (A) tail signals, AATAAA sequence was at site 65 base upstream from the 3' terminus. The deduced amino acid sequence of p10 protein was 95 with a predicted molecular weight of 10.26 kDa. In the p10 protein sequence, a hydrophobic region was present at the N-terminus of the protein, whereas the C-terminus was highly hydrophilic. The p10 protein of H. cunea NPV did not contain cysteine, histidine, trytophan, tryptophane, tyrosine, glutamine and asparagine residues.

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Nucleotide Sequence of Rat Transketolase and Liver-Specific Pretranslational Activation During Postnatal Development

  • Kim, Sung-Min F.;Kim, Byung-Moon;Jeng, Jingjau;Soh, Yun-Jo;Bak, Choong-Il;Huh, Jae-Wook;Song, Byoung-J.
    • BMB Reports
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    • v.29 no.2
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    • pp.146-150
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    • 1996
  • A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

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Intrinsic bent DNA colocalizes with the sequence involved in the Nd-sD mutation in the Bombyx mori fibroin light chain gene

  • Barbosa, Joice Felipes;Bravo, Juliana Pereira;Takeda, Karen Izumi;Zanatta, Daniela Bertolini;Silva, Jose Luis Da Conceicao;Balani, Valerio Americo;Fiorini, Adriana;Fernandez, Maria Aparecida
    • BMB Reports
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    • v.41 no.5
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    • pp.394-399
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    • 2008
  • Multiple sequence alignments of the Bombyx mori fibroin light chain gene (fib-L) from hybrids and from Chinese and Japanese strains demonstrated that 51.6% of the fib-L third intron is conserved. One of these conserved segments, 41 bp long, contains the sequence CGTTATTATACATATT, which is duplicated in the B. mori Nd-$s^D$ mutant. In the present work, electrophoretic mobility assays and computational analyses revealed a major peak of intrinsic bent DNA within the segment that undergoes breakage in the previously-described Nd-$s^D$ mutation. This result suggested that this intrinsically-curved region might mediate DNA cleavage and enhance recombination events in the third intron of the Bombyx mori fib-L gene.

Mitochondrial sequence based characterization and morphometric assessment of Diara buffalo population

  • Singh, Karan Veer;Purohit, Hitesh;Singh, Ramesh Kumar
    • Animal Bioscience
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    • v.35 no.7
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    • pp.949-954
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    • 2022
  • Objective: The present study is aimed at phenotypic characterization and mitochondrial d-loop analysis of indigenous "Diara" buffalo population, which are mostly confined to the villages on the South and North Gangetic marshy plains in the Bihar state of India. These buffaloes are well adapted and are best suited for ploughing and puddling the wet fields meant for paddy cultivation. Methods: Biometric data on 172 buffaloes were collected using a standard flexible tape measure. Animals are medium in size; the typical morphometric features are long head with a broad forehead and moderately long and erect ears. Genomic DNA was isolated from unrelated animals. The mtDNA d-loop 358-bp sequence data was generated and compared with 338 sequences belonging to riverine and swamp buffaloes. Results: Based on the mitochondrial d-loop analysis the Diara buffaloes were grouped along with the haplotypes reported for riverine buffalo. Sequence analysis revealed the presence of 7 mitochondrial D loop haplotypes with haplotype diversity of 0.9643. Five of the haplotypes were shared with established swamp breeds and with Buffalo population of Orissa in India. Conclusion: Morphometric analyses clearly shows distinguishing features like long and broad forehead which may be useful in identification. The germplasm of Diara buffalo is much adapted to the marshy banks of river Ganga and its tributaries. It constitutes a valuable genetic resource which needs to be conserved on priority basis.

Next-generation sequencing for the genetic characterization of Maedi/Visna virus isolated from the northwest of China

  • Zhao, Ling;Zhang, Liang;Shi, Xiaona;Duan, Xujie;Li, Huiping;Liu, Shuying
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.66.1-66.9
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    • 2021
  • Background: Maedi/Visna virus (MVV) is a contagious viral pathogen that causes considerable economic losses to the sheep industry worldwide. Objectives: In China, MVV has been detected in several regions, but its molecular characteristics and genetic variations were not thoroughly investigated. Methods: Therefore, in this study, we conducted next-generation sequencing on an MVV strain obtained from northwest China to reveal its genetic evolution via phylogenetic analysis. Results: A MVV strain obtained from Inner Mongolia (NM) of China was identified. Sequence analysis indicated that its whole-genome length is 9193 bp. Homology comparison of nucleotides between the NM strain and reference strains showed that the sequence homology of gag and env were 77.1%-86.8% and 67.7%-75.5%, respectively. Phylogenetic analysis revealed that the NM strain was closely related to the reference strains isolated from America, which belong to the A2 type. Notably, there were 5 amino acid insertions in variable region 4 and a highly variable motif at the C-terminal of the surface glycoprotein (SU5). Conclusions: The present study is the first to show the whole-genome sequence of an MVV obtained from China. The detailed analyses provide essential information for understanding the genetic characteristics of MVV, and the results enrich the MVV library.

Unraveling Haplotype Diversity of the Apical Membrane Antigen-1 Gene in Plasmodium falciparum Populations in Thailand

  • Lumkul, Lalita;Sawaswong, Vorthon;Simpalipan, Phumin;Kaewthamasorn, Morakot;Harnyuttanakorn, Pongchai;Pattaradilokrat, Sittiporn
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.153-165
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    • 2018
  • Development of an effective vaccine is critically needed for the prevention of malaria. One of the key antigens for malaria vaccines is the apical membrane antigen 1 (AMA-1) of the human malaria parasite Plasmodium falciparum, the surface protein for erythrocyte invasion of the parasite. The gene encoding AMA-1 has been sequenced from populations of P. falciparum worldwide, but the haplotype diversity of the gene in P. falciparum populations in the Greater Mekong Subregion (GMS), including Thailand, remains to be characterized. In the present study, the AMA-1 gene was PCR amplified and sequenced from the genomic DNA of 65 P. falciparum isolates from 5 endemic areas in Thailand. The nearly full-length 1,848 nucleotide sequence of AMA-1 was subjected to molecular analyses, including nucleotide sequence diversity, haplotype diversity and deduced amino acid sequence diversity and neutrality tests. Phylogenetic analysis and pair-wise population differentiation ($F_{st}$ indices) were performed to infer the population structure. The analyses identified 60 single nucleotide polymorphic loci, predominately located in domain I of AMA-1. A total of 31 unique AMA-1 haplotypes were identified, which included 11 novel ones. The phylogenetic tree of the AMA-1 haplotypes revealed multiple clades of AMA-1, each of which contained parasites of multiple geographical origins, consistent with the $F_{st}$ indices indicating genetic homogeneity or gene flow among geographically distinct populations of P. falciparum in Thailand's borders with Myanmar, Laos and Cambodia. In summary, the study revealed novel haplotypes and population structure needed for the further advancement of AMA-1-based malaria vaccines in the GMS.