• Title/Summary/Keyword: Molecular phylogenetic analysis

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Distribution and genetic diversity of Feline calicivirus in Moscow metropolitan area

  • Alina, Komina;Nikita, Krasnikov;Oksana, Kucheruk;Elena, Zhukova;Anton, Yuzhakov;Alexey, Gulyukin
    • Journal of Veterinary Science
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    • v.23 no.6
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    • pp.92.1-92.8
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    • 2022
  • Background: Feline calicivirus (FCV) is widespread throughout the world. An FCV infection is associated with conjunctivitis, rhinitis, and mouth ulcers that can lead to the animal's death. Because vaccination is not always effective, it is necessary to monitor the infection regularly. Objectives: This study examined the FCV epizootic situation in the Moscow metropolitan area by conducting a molecular phylogenetic analysis of the virus isolates. Methods: Samples from 6213 animals were examined by a reverse transcription polymerase chain reaction. For phylogenetic analysis, 12 nucleotide sequences obtained from animal samples were selected. Sequencing was performed using the Sanger method. Phylogenetic analysis was conducted using the Maximum Likelihood method. Results: The FCV genome was detected in 1,596 (25.7%) samples out of 6,213. In 2018, calicivirus was detected in 18.9% of samples, 27.8% in 2019, 21.4% in 2020, and 32.6% in 2021. Phylogenetic analysis of the F ORF2 region and the ORF3 start region led to division into two FCV genogroups. Most of the isolates (8 out of 12) were close to the Chinese strains. On the other hand, there were isolates closely related to European and American strains. The isolates circulating in Moscow were not included in clusters with vaccine strains; their nucleotide similarity varied from 77% to 83%. Conclusions: This study revealed a high prevalence and genetic diversity of the FCV in Moscow. The epizootic situation remains stably tense because 24 viruses were detected in 25% of animals annually.

Cloning of Acetate Kinase Gene from the Copepod Paracyclopina nana and its Expression in Escherichia coli (요각류 Paracyclopina nana Acetate Kinase의 클로닝 및 대장균에서의 발현)

  • Jung Sang-Oun;Seo Jung Soo;Lee Young-Mi;Park Tae-Jin;Kim Il-Chan;Park Heum Gi;Lee Jae-Seong
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.157-163
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    • 2005
  • The acetate kinase gene from the copepod Paracyclopina nana was cloned. The open reading frame (ORF) was 1,200 bp, and poly(A) signal sequence was located in the end of the ORF. After the molecular phylogenetic analysis of P nana acetate kinase gene, it was revealed that it formed the same branch with that of Aspergillus. Also P. nana acetate kinase showed the difference with those of other prokaryotic microorganisms but showed the same clade with those of fungi. We also confirmed that the recombinant protein of P. nana acetate kinase made approximately 50 kDa after expression of recombinant gene construct in E. coli. This may be useful to compare this protein to those of other organisms in biochemical characteristics.

Characterization of Ty3-gypsy-like Elements in Hibiscus syriacus

  • Jeung, Ji-Ung;Cho, Sung Ki;Lee, Seung Jae;Shin, Jeong Sheop
    • Molecules and Cells
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    • v.19 no.3
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    • pp.318-327
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    • 2005
  • Southern blot analysis revealed a ubiquitous distribution and high copy number of Ty3-gypsy-like elements in the genome of Hibiscus syriacus. Comparative phylogenetic analysis of the large subunit of Rubisco and the integrase region of Ty3-gypsy elements in various plant species indicated that the retrotransposon-like sequences have different evolutionary histories and their own unique polymorphism in the H. syriacus population. Sequence-tagged site-restriction fragment length polymorphisms (STS-RFLP) analysis also indicated great variability in the numbers and sequences of Ty3-gypsy-like elements within H. syriacus varieties. Ty3-gypsy-like elements may still be active within H. syriacus, since Northern analysis of wounded leaves of H. syriacus variety Saehan with a probe for the integrase domain gave strong hybridization signals. The sequence heterogeneity and ubiquity of the Ty3-gypsy-like elements in H. syriacus genomes could provide reliable DNA markers for line identification as well for the analysis of genetic diversity in H. syriacus.

Phylogeny of Ganoderma Based on the Restriction Enzyme Analysis of Mitochondrial DNA (미토콘드리아 DNA의 제한효소 분석법에 의한 영지의 계통분류)

  • Hong, Soon-Gyu;Jung, Hack-Sung
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.245-251
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    • 1994
  • Ten strains of 7 species from the genus Ganoderma, G. lucidum ATCC 64251, FP-103561-T, and ES70701, G. applanatum ATCC 44053 and FP-57035-T. G. lobatum ATCC 42985, G. resinaceum ATCC 52416, G. subamboinense var. laevisporum ATCC 52420, G. meredithae ATCC 64492, and G. microsporum ATCC 76024, were studied to discuss their phylogenetic relationships by utilizing restriction fragment length polymorphisms (RFLPs) of mitochondrial DNAs (mtDNAs). Six restriction enzymes, BamHI, BglII, EcoRI, HindIII, PvuII, and XbaI which digested mtDNAs into adequate numbers of restriction fragments for cluster analysis, were used in this study. Restriction profiles of strains for each restriction enzyme were treated as analysis characters to calculate similarity coefficients, which were converted into nucleotide sequence divergence values whose mean values were then arranged in a matrix table. This table was utilized for a phylogenetic analysis using the Neighborjoining method of the PHYLIP package to construct phylogenetic tree. Three strains of G. lucidum and two strains of G. applanatum exhibited different lineages each but one of G. applanatum strains showed a close relationship with G. lobatum, which reflected the species complexity of these species whose strains were phenotypically indistinguishable but genetically distinct. The present results suggest that the natural classification of Ganoderma needs to be considered from the viewpoints of molecular biology-based systematics as well as morphological classifications and cultural identifications for better phylogenetic conclusions.

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Mitochondrial DNA variation and phylogeography of native Mongolian goats

  • Ganbold, Onolragchaa;Lee, Seung-Hwan;Paek, Woon Kee;Munkhbayar, Munkhbaatar;Seo, Dongwon;Manjula, Prabuddha;Khujuu, Tamir;Purevee, Erdenetushig;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.902-912
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    • 2020
  • Objective: Mongolia is one of a few countries that supports over 25 million goats, but genetic diversity, demographic history, and the origin of goat populations in Mongolia have not been well studied. This study was conducted to assess the genetic diversity, phylogenetic status and population structure of Mongolian native goats, as well as to discuss their origin together with other foreign breeds from different countries using hypervariable region 1 (HV1) in mtDNA. Methods: In this study, we examined the genetic diversity and phylogenetic status of Mongolian native goat populations using a 452 base-pair long fragment of HVI of mitochondrial DNA from 174 individuals representing 12 populations. In addition, 329 previously published reference sequences from different regions were included in our phylogenetic analyses. Results: Investigated native Mongolian goats displayed relatively high genetic diversities. After sequencing, we found a total of 109 polymorphic sites that defined 137 haplotypes among investigated populations. Of these, haplotype and nucleotide diversities of Mongolian goats were calculated as 0.997±0.001 and 0.0283±0.002, respectively. These haplotypes clearly clustered into four haplogroups (A, B, C, and D), with the predominance of haplogroup A (90.8%). Estimates of pairwise differences (Fst) and the analysis of molecular variance values among goat populations in Mongolia showed low genetic differentiation and weak geographical structure. In addition, Kazakh, Chinese (from Huanghuai and Leizhou), and Arabian (Turkish and Baladi breeds) goats had smaller genetic differentiation compared to Mongolian goats. Conclusion: In summary, we report novel information regarding genetic diversity, population structure, and origin of Mongolian goats. The findings obtained from this study reveal that abundant haplogroups (A to D) occur in goat populations in Mongolia, with high levels of haplotype and nucleotide diversity.

Analysis of the Bioactive Metabolites of the Endangered Mexican Lost Fungi Campanophyllum - A Report from India

  • Borthakur, Madhusmita;Gurung, Arun Bahadur;Bhattacharjee, Atanu;Joshi, S.R.
    • Mycobiology
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    • v.48 no.1
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    • pp.58-69
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    • 2020
  • Meghalaya, (in India), in the region of the mega-biodiversity hotspots, is home to a plethora of wild mushrooms. The present study concerns the exploration of the order Agaricales, which includes rare gilled mushrooms considered endangered under IUCN A4c criteria, due to the declining habitat. Electron microscopy of the gill sections revealed an abundance of clamp connections, hyphal cell walls, cystidia, and basidia. This rare species which belongs to the family Cyphellaceae, exhibits morphological and molecular differences from the Cyphella spp. Phylogenetic analysis revealed that it formed a clade under the genus Campanophyllum of the order Agaricales, confirmed by both Neighbor Joining (NJ) and Bayesian phylogenetic analysis. Being nutritionally potent along with its efficient antioxidant value, the fungal extract shows significant rise of two-fold in the antimicrobial activity along with the commercial antibiotics. The compound, Phenol, 2, 4-bis (1, 1-Dimethylethyl) (2, 4-DTBP) showed in ample range in the fungal extract along with aliphatic hydrocarbons, terpene, alcohol and volatile organic compounds on further characterization in GCMS. The present study indicates the endangered Campanophyllum proboscideum could be a rich source of natural antioxidants and an effective pharmaceutical agent.

Molecular Phylogeny of the Gayal in Yunnan China Inferred from the Analysis of Cytochrome b Gene Entire Sequences

  • Li, S.P.;Chang, H.;Ma, G.L.;Cheng, H.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.6
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    • pp.789-793
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    • 2008
  • The gayal (Bos frontalis) in China is a very rare semi-wild and semi-domestic bovine species. There still exist remarkable divergences on the gayal's origin and taxonomic status. In the present study, the cytochrome b (Cyt b) gene entire sequences (1,140 bp) of 11 gayals in Yunnan China were analyzed. Combined with other bovine Cyt b sequences cited in GenBank, the phylogenetic trees of genus Bos were reconstructed by neighbor-joining (NJ) and maximum parsimony (MP) methods with Bubalus bubalis as outgroup. Sequence analysis showed that, among 1,140 sites compared for 11 gayals, 95 variable sites (8.33% of all sites) and 6 different haplotypes were observed, showing abundant mitochondrial genetic diversity in gayals. Both NJ and MP trees demonstrated that gayals in this study were markedly divided into three embranchments: one embranchment clustering with Bos gaurus, another clustering with Bos taurus, and the third clustering with Bos indicus. The result of phylogenetic analysis suggested that the gayal might be the domesticated form of the gaur, and a great proportion of the gayal bloodline in China was invaded by other bovine species.

Prevalence and molecular characterization of tetracycline-resistant Enterococcus isolates from livestock (가축에서 테트라사이클린 내성 장구균 조사 및 분자생물학적 특성규명)

  • Kim, Chul-Min;Kang, Su-Jin;Lee, Beyong-Jong;Lee, Sung-Jae;Yuk, Dae-Su
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.143-149
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    • 2010
  • BIn the present study, Enterococcus isolates originating from livestock were studied for the phenotypic and genotypic assessment of tetracycline resistance. A total of 74 isolates encompassing the species Enterococcus faecalis (n=12) and E. faecium (n=62) displayed phenotypic resistance to tetracycline. Tetracycline resistance gene [tet (M), 1,886bp] were sequenced by dye terminator cycle sequencing method and compared with tet (M) sequences available from the GenBank database. Sequencing analysis of PCR amplicons showed high homology to the reference strains ranging 97.2~100%. The tet (M) genes were divided into three major subgroups according to phylogenetic analysis. The genetic information obtained from this study could be useful for the molecular study of enterococci.

Genetic Variation of Rice Populations Estimated Using nrDNA ITS Region Sequence

  • Wang, Dong;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.249-255
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    • 2014
  • The rice belonging to Oryza sativa is not only has significant economic importance, for it is the major source of nutrition for about 3 billion all around the world. But also plays a vital role as a model organism, because it has a number of advantages to be a model plant, such as efficient transformation system and small genome size. Many methods and techniques have been conducted to attempt to distinguish different Oryza sativa species, such as amplified fragment length polymorphism (AFLP), random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and so on. However, studies using sequence analysis of internal transcribed spacer (ITS), a region of ribosomal RNA has not been reported until now. This study was undertaken with an aim to understand the phylogenetic relationships among sixteen isolates of Oryza sativa collected from abroad and fifteen isolates collected from Korea, using ribosomal RNA (rRNA) internal transcribed spacer (ITS) sequences to compare the phylogeny relationships among different Oryza sativa species. The size variation obtained among sequenced nuclear ribosomal DNA (nrDNA) ITS region ranged from 515bp to 1000bp. The highest interspecific genetic distance (GD) was found between Sfejare 45 (FR12) and Anapuruna (FR15). Taebong isolate showed the least dissimilarity of the ITS region sequence with other thirty isolates. This consequence will help us further understanding molecular diversification in intra-species population and their phylogenetic analysis.

Molecular Phylogeny of the Family Strigidae (Aves) Based on Mitochondrial Cytochrome b Gene (Mitochondrial Cytochrome b 유전자에 의한 올빼미과 (Family Strigidae)의 분자계통)

  • 류시현;박희천
    • Animal Systematics, Evolution and Diversity
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    • v.19 no.2
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    • pp.297-304
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    • 2003
  • Phylogenetic analysis of 31 species representing 12 genera in the family Strigidae (Aves: Strigiformes) including 5 species (Bubo bubo, Otus sunia, O. semitorques, Ninox scutulato, Strix aluco) collected from Korea has been undertaken using nucleotide sequences of the mitochondrial cytochrome b gene. Maximum likelihood analysis was performed and pairwise genetic distances were calculated with Kimura's two-parameter and p-distance. Among well-aligned 959 bp used for this study, 459 sites were variable and 398 sites were informative for the phylogenetic analysis. The family Strigidae was divided into three subgroups, Clade I (Aegolius), Clade II (Athene, Micrathene, Glaucidium and Surnia) and Clade III (Bubo, Nycteo, Pulsatrix, Strix, Otus, Ptilopsis, and Ninox). Also, two separated subgroups in the genus Otus were confirmed by the geographical distribution.