• Title/Summary/Keyword: Sequence-based molecular analysis

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Molecular Phylogenetic study of Acila divaricata vigila based on the Partial Sequence of 16S rRNA Gene (민호두조개 (Acila divaricata vigila) 의 16S rRNA 유전자를 기초로 한 분자계통 분류학적 연구)

  • Kim, Bong-Seok;Kang, Se-Won;Jeong, Ji-Eun;Park, Jung-Yeon;Kang, Jung-Ha;Han, Yeon-Soo;Ko, Hyun-Sook;An, Chel-Min;Lee, Jun-Sang;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.395-400
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    • 2011
  • Phylogenetic analyses on the Phylum Mollusks has so far been conducted by many researchers in the world. However, there was no report on taxonomic analysis on Acila divaricata vigila which is belonging to Class Bivalvia, Subclass Protobranchia. In this study, we performed molecular phylogenetic analysis on Acila divaricata vigila using 16S rRNA sequence through maximum likelihood method. As a result, it is clearly divided into the legion of mollusk classification unit (when you zoom in order) and represented to support the current classification in the Phylum Mollusca belong to Class Bivalvia, Subclass Protobranchia, Subclass Pteriomorphia, Subclass Paleoheterodonta, Subclass Heterodonta and Subclass Anomalodesmacea. To our knowledge, this is the first report of molecular phylogenetic analysis on Acila divaricata vigila using 16S rRNA gene and these data suggests that 16S rRNA gene will be useful for analyzing the phylogenetic relationship of Subclass Protobranchia.

Molecular Identification of Diphyllobothrium nihonkaiense from 3 Human Cases in Heilongjiang Province with a Brief Literature Review in China

  • Zhang, Weizhe;Che, Fei;Tian, Song;Shu, Jing;Zhang, Xiaoli
    • Parasites, Hosts and Diseases
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    • v.53 no.6
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    • pp.683-688
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    • 2015
  • Human diphyllobothriasis is a widespread fish-borne zoonosis caused by the infection with broad tapeworms belonging to the genus Diphyllobothrium. In mainland China, so far 20 human cases of Diphyllobothrium infections have been reported, and the etiologic species were identified as D. latum and D. nihonkaiense based on morphological characteristics or molecular analysis. In the present study, proglottids of diphyllobothriid tapeworms from 3 human cases that occurred in Heilongjiang Province, China were identified as D. nihonkaiense by sequencing mitochondrial cytochrome c oxidase subunit I (cox1) and NADH dehydrogenase subunit 5 (nad5) genes. Two different cox1 gene sequences were obtained. One sequence showed 100% homology with those from humans in Japan. The remaining cox1 gene sequence and 2 different nad5 gene sequences obtained were not described previously, and might reflect endemic genetic characterizations. D. nihonkaiense might also be a major causative species of human diphyllobothriasis in China. Meanwhile, the finding of the first pediatric case of D. nihonkaiense infection in China suggests that infants infected with D. nihonkaiense should not be ignored.

Mitochondrial DNA Sequence Variations and Genetic Relationships among Korean Thais Species (Muricidae: Gastropoda)

  • Lee, Sang-Hwa;Kim, Tae-Ho;Lee, Jun-Hee;Lee, Jong-Rak;Park, Joong-Ki
    • Animal Systematics, Evolution and Diversity
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    • v.27 no.1
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    • pp.1-17
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    • 2011
  • Thais Roding, 1798, commonly known as rock-shell, is among the most frequently found gastropod genera worldwide on intertidal rocky shores including those of Japan, China, Taiwan and Korea. This group contains important species in many marine environmental studies but species-level taxonomy of the group is quite complicated due to the morphological variations in shell characters. This study examined the genetic variations and relationships among three Korean Thais species based on the partial nucleotide sequences of mitochondrial cox1 gene fragments. Phylogenetic trees from different analytic methods (maximum parsimony, neighbor-joining, and maximum likelihood) showed that T. bronni and T. luteostoma are closely related, indicating the most recent common ancestry. The low sequence divergence found between T. luteostoma and T. bronni, ranging from 1.53% to 3.19%, also corroborates this idea. Further molecular survey using different molecular marker is required to fully understand a detailed picture of the origin for their low level of interspecific sequence divergence. Sequence comparisons among conspecific individuals revealed extensive sequence variations within the three species with maximum values of 2.43% in T. clavigera and 1.37% in both T. bronni and T. luteostoma. In addition, there is an unexpectedly high level of mitochondrial genotypic diversity within each of the three Korean Thais species. The high genetic diversity revealed in Korean Thais species is likely to reflect genetic diversity introduced from potential source populations with diverse geographic origins, such as Taiwan, Hong Kong, and a variety of different coastal regions in South China and Japan. Additional sequence analysis with comprehensive taxon sampling from unstudied potential source populations will be also needed to address the origin and key factors for the high level of genetic diversity discovered within the three Korean Thais species studied.

Characterization of Sclerotinia sclerotiorum Isolated from Paprika

  • Jeon, Young-Jae;Kwon, Hyuk-Woo;Nam, Ji-Sun;Kim, Seong-Hwan
    • Mycobiology
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    • v.34 no.3
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    • pp.154-157
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    • 2006
  • A fungal isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Sclerotinia sclerotiorum based on its ability of sclerotium formation, physiological and molecular properties. When the isolate was grown on potato dextrose agar, oatmeal agar, and malt extract agar, it grew most well on PDA. Optimal temperature and pH for its growth were $25^{\circ}C$ and pH 7, respectively. The fungal isolate produced sclerotia on PDA within 10 days, and the color and shape of the sclerotia were similar to those of S. sclerotiorum. The ITS rDNA regions including ITS1 and ITS2 and 5.8S sequences were amplified using ITS1F and ITS4 primers from the genomic DNAs of the paprika isolate and other known pathogenic S. sclerotiorum isolated from different crops in Korea, and their nucleotide sequences were determined. Sequence comparison analysis showed the ITS rDNA of the paprika isolate shares 100% sequence identity with those of S. sclerotiorum isolated from red pepper, lettuce and a S. sclerotiorum isolate registered in GenBank DNA database. Neighbor joining analysis based on the ITS rDNA sequence revealed the paprika isolate has very close phylogenetic relationships with known Sclerotinia sclerotiorum isolates. This is the first report that S. sclerotiorum has been found associated with paprika rot in paprika growing countries.

Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF)- Based Cloning of Enolase, ENO1, from Cryphonectria parasitica

  • Kim, Myoung-Ju;Chung, Hea-Jong;Park, Seung-Moon;Park, Sung-Goo;Chung, Dae-Kyun;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.620-627
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    • 2004
  • On the foundation of a database of genome sequences and protein analyses, the ability to clone a gene based on a peptide analysis is becoming more feasible and effective for identifying a specific gene and its protein product of interest. As such, the current study conducted a protein analysis using 2-D PAGE followed by MALDI- TOF and ESI-MS to identify a highly expressed gene product of C. parasitica. A distinctive and highly expressed protein spot with a molecular size of 47.2 kDa was randomly selected and MALDI-TOF MS analysis was conducted. A homology search indicated that the protein appeared to be a fungal enolase (enol). Meanwhile, multiple alignments of fungal enolases revealed a conserved amino acid sequence, from which degenerated primers were designed. A screening of the genomic $\lambda$ library of C. parasitica, using the PCR amplicon as a probe, was conducted to obtain the full-length gene, while RT-PCR was performed for the cDNA. The E. coli-expressed eno 1 exhibited enolase enzymatic activity, indicating that the cloned gene encoded the C. parasitica enolase. Moreover, ESI-MS of two of the separated peptides resolved from the protein spot on 2-D PAGE revealed sequences identical to the deduced sequences, suggesting that the cloned gene indeed encoded the resolved protein spot. Northern blot analysis indicated a consistent accumulation of an eno1 transcript during the cultivation.

Analysis of Molecular Variance and Population Structure of Sesame (Sesamum indicum L.) Genotypes Using Simple Sequence Repeat Markers

  • Asekova, Sovetgul;Kulkarni, Krishnanand P.;Oh, Ki Won;Lee, Myung-Hee;Oh, Eunyoung;Kim, Jung-In;Yeo, Un-Sang;Pae, Suk-Bok;Ha, Tae Joung;Kim, Sung Up
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.321-336
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    • 2018
  • Sesame (Sesamum indicum L.) is an important oilseed crop grown in tropical and subtropical areas. The objective of this study was to investigate the genetic relationships among 129 sesame landraces and cultivars using simple sequence repeat (SSR) markers. Out of 70 SSRs, 23 were found to be informative and produced 157 alleles. The number of alleles per locus ranged from 3 - 14, whereas polymorphic information content ranged from 0.33 - 0.86. A distance-based phylogenetic analysis revealed two major and six minor clusters. The population structure analysis using a Bayesian model-based program in STRUCTURE 2.3.4 divided 129 sesame accessions into three major populations (K = 3). Based on pairwise comparison estimates, Pop1 was observed to be genetically close to Pop2 with $F_{ST}$ value of 0.15, while Pop2 and Pop3 were genetically closest with $F_{ST}$ value of 0.08. Analysis of molecular variance revealed a high percentage of variability among individuals within populations (85.84%) than among the populations (14.16%). Similarly, a high variance was observed among the individuals within the country of origins (90.45%) than between the countries of origins. The grouping of genotypes in clusters was not related to their geographic origin indicating considerable gene flow among sesame genotypes across the selected geographic regions. The SSR markers used in the present study were able to distinguish closely linked sesame genotypes, thereby showing their usefulness in assessing the potentially important source of genetic variation. These markers can be used for future sesame varietal classification, conservation, and other breeding purposes.

Sequence Analysis of $\beta$-Xylosidase Gene from Bacillus stearothermophilus (Bacillus stearothermophilus $\beta$-Xylosidase 유전자의 염기 서열 결정 및 분석)

  • 오현주;최용진
    • Microbiology and Biotechnology Letters
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    • v.22 no.2
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    • pp.134-142
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    • 1994
  • The neucleotide sequences of the xylA gene encoding $\beta $-xylosidase of Bacillus stearothermophilus and is its flanking regions were datermined. Three open reading frame(ORFs) were found, one of which(ORF1) appeared to code for the $\beta $-xylosidase. The 1830 base pair ORF1 encoded 609 amino acids starting from a TTG initiation codon. The molecular weight deduced from the nucleotide sequence(68 KD) was in agreement with that estimated by SDS-polyacrylamide gel electrophoresis of the purified enzyme(66 KD). The Shine-Dalgarno sequence(5'-AGGAGG-3') was found 11 bp upstream of the initiation codon. Further 15 bp upstream, there observed a potential transcription initiation signals. The putative -10 sequence(CATAAT) and -35 sequence(TTGTTA) coresponded closely to the consensus sequences for Bacillus subtilis RNA polymerase with major sigma factor. The guanine-plus-cytosine content of the coding region of the xylA gene was 56mol% while that of the third position of the codons was 63 mol%. Based on the comparison with the amino acid sequences of several other carbohydrate degrading enzymes, two conserved regions, possibly participating in the catalytic mechamism of $\beta $-xylosidase xylA, were identified in 278-298 and 329-350 regions of the translated xylA gene. The nucleotide sequence of the xylA was found to exhibit no homology to any other genes so far reproted.

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Sequence Validation for the Identification of the White-Rot Fungi Bjerkandera in Public Sequence Databases

  • Jung, Paul Eunil;Fong, Jonathan J.;Park, Myung Soo;Oh, Seung-Yoon;Kim, Changmu;Lim, Young Woon
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1301-1307
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    • 2014
  • White-rot fungi of the genus Bjerkandera are cosmopolitan and have shown potential for industrial application and bioremediation. When distinguishing morphological characters are no longer present (e.g., cultures or dried specimen fragments), characterizing true sequences of Bjerkandera is crucial for accurate identification and application of the species. To build a framework for molecular identification of Bjerkandera, we carefully identified specimens of B. adusta and B. fumosa from Korea based on morphological characters, followed by sequencing the internal transcribed spacer region and 28S nuclear ribosomal large subunit. The phylogenetic analysis of Korean Bjerkandera specimens showed clear genetic differentiation between the two species. Using this phylogeny as a framework, we examined the identification accuracy of sequences available in GenBank. Analyses revealed that many Bjerkandera sequences in the database are either misidentified or unidentified. This study provides robust reference sequences for sequence-based identification of Bjerkandera, and further demonstrates the presence and dangers of incorrect sequences in GenBank.

Molecular Cloning of Peroxidase cDNA from Flower Bud of Ginseng(Panax ginseng)

  • In Jun-Gyo;Lee Bum-Soo;Song Won-Seob;Bae Chang-Hyu;Choi Seong-Kyu;Yang Deok-Chun
    • Plant Resources
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    • v.8 no.2
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    • pp.155-160
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    • 2005
  • A full-length cDNA (PPrx1) encoding peroxidase has been isolated and its nucleotide sequence determined from flower bud in ginseng plant (Panax ginseng). A PPrx1 cDNA is 1192 nucleotides long and has an open reading frame of 1062 bp with a deduced amino acid sequence of 354 residues (pI 7.53). The deduced amino acid sequence of PPrx1 matched to the previously reported peroxidase protein genes. The PPrx1 showed a high similarity with the $64\%$ identity with peroxidase of N. tabacum (AAK52084). In the phylogenetic analysis based on the amino acid residues, the PPrx1 was closer with peroxidase of G. max (AAD37376).

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Molecular Characterization of Plasmid from Bifidobacterium longum

  • Park, Myeong-Soo;Moon, Hye-Won;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.457-462
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    • 2003
  • The complete nucleotide sequence of a plasmid, pMG1, isolated from Bifidobacterium longum MG1 has been determined. This plasmid, composed of 3,862 base pairs with 65.1% of G+C content. harbors two major open reading frames (ORF) encoding putative proteins of 29 kDa (ORF I) and 71 kDa (ORF II). ORF I showed relatively high amino acid sequence homology with replication proteins of other plasmids from Gr Im-positive and -negative bacteria. Upstream of ORF I, four sets of tandem repeat sequences resembling the iteron structure of related plasmids were found. S1 endonuclease treatment and Southern blot analysis revealed that pMG1 accumulates single-stranded DNA (ssDNA) intermediate, which indicate i the rolling circle replication (RCR) mechanism of this plasmid. Homology search indicated that ORF II encodes plasmid mobilization protein, and the presence of highly conserved oriT sequence in the upstream of this gene supported this assumption. RT-PCR showed that only ORF I is expressed in vivo. Based on these results, pMG 1 was exploited to construct a shuttle vector, pBES2. It was successfully transformed into Bifidobacterium and maintained stably.