• Title/Summary/Keyword: Sequence Analysis, DNA

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Identification of Bacteria Causing Fermentation of Oriental Melon in Korea (참외 발효과를 유발하는 세균의 동정)

  • Choi, Jae-Eul;Cha, Sun-Kyung;Kim, Jin-Hee;Yuk, Jin-Ah;Hwang, Yong-Soo;Kwon, Soon-Wo
    • Research in Plant Disease
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    • v.9 no.4
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    • pp.189-195
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    • 2003
  • Bacteria causing fermentation in Oriental melon were identified as three independent groups on the basis of 16S rDNA sequence analysis. The 16S rDNA sequence of the strain CM2105 showed the highest identity (99.6%) with that of Microbacterium phyllosphaerae, and also indicated high sequence identity to that of M. holiorum (99.5%). The 16S rDNA sequences of the strain CM2101 and CM2121 matched at the high sequence similarity (98.9%, 98.8, respectively), to that of Pseudomonas pavonacea, and the DNA sequence of CM2126 showed high sequence identity to that of P. costantinii (99.5%), and P. grimontii (99.0%). The 16S rDNA sequence of the strain CM2113 showed the highest identity (99.7%) with that of Enterobacter cloacae. The 16S rDNA sequences, the physiological and biochemical analysis suggested that the strain CM2105 belonged to Microbacterium phyllosphaerae, CM2101, CM2121 and CM2126 to Pseudomonas spp., CM2113 to Enterobacter cloacae.

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.

Phylogeny of Flavobacteria Group Isolated from Freshwater Using Multilocus Sequencing Analysis

  • Mun, Seyoung;Lee, Jungnam;Lee, Siwon;Han, Kyudong;Ahn, Tae-Young
    • Genomics & Informatics
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    • v.11 no.4
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    • pp.272-276
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    • 2013
  • Sequence analysis of the 16S rRNA gene has been widely used for the classification of microorganisms. However, we have been unable to clearly identify five Flavobacterium species isolated from a freshwater by using the gene as a single marker, because the evolutionary history is incomplete and the pace of DNA substitutions is relatively rapid in the bacteria. In this study, we tried to classify Flavobacterium species through multilocus sequence analysis (MLSA), which is a practical and reliable technique for the identification or classification of bacteria. The five Flavobacterium species isolated from freshwater and 37 other strains were classified based on six housekeeping genes: gyrB, dnaK, tuf, murG, atpA, and glyA. The genes were amplified by PCR and subjected to DNA sequencing. Based on the combined DNA sequence (4,412 bp) of the six housekeeping genes, we analyzed the phylogenetic relationship among the Flavobacterium species. The results indicated that MLSA, based on the six housekeeping genes, is a trustworthy method for the identification of closely related Flavobacterium species.

Prediction of Rice Embryo Proteins using EST-Databases

  • Woo, Sun-Hee;Cho, Seung-Woo;Kim, Tae-Seon;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Jong, Seung-Keun
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.1-7
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    • 2008
  • An attempt was made to link rice embryo proteins to DNA sequences and to understand their functions. One hundred of the 700 spots detected on the embryo 2-DE gels were microsequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were unknown in functions as previously reported (Woo et al. 2002). In addition, twenty-four protein spots with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available rice EST databases at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). The chromosomal location of some proteins were also obtained from the rice genetic map provided by Japanese Rice Genome Research Program (http://rgp.dna.affrc.go.jp). The DNA sequence databases including EST have been reported for rice (Oryza sativa L.) now provides whole or partial gene sequence, and recent advances in protein characterization allow the linking proteins to DNA sequences in the functional analysis. This work shows that proteome analysis could be a useful tool strategy to link sequence information and to functional genomics.

cDNA Sequence and mRNA Expression of a Novel Peroxiredoxin from the Firefly, pyrocoelia rufa

  • Jin, Byung-Rae;Lee, Kwang-Sik;Kim, Seong-Ryul;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.2
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    • pp.101-107
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    • 2002
  • We describe here the cDNA sequence and mRNA expression of a novel family of the antioxidant protein, peroxiredoxin, from the firefly, Pyracoetia ruin. The 555 bp cDNA sequence codes for a 185 amino acid protein with a calculated molecular mass of approximately 21 kDa. The deduced protein of P. rufa peroxiredoxin gene contains two conserved cysteine residues. Alignment of the deduced protein of P. rufa peroxiredoxin gene showed 71.1% protein sequenceidentity to known insect Drosophila melanogaster peroxiredoxin. Northern blot analysis revealed that the P. rufa peroxiredoxin is specifically expressed in the fat body of P. rufa larvae.

DNA Chip Technologies

  • Hwang, Seoung-Yong;Lim, Geun-Bae
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.159-163
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    • 2000
  • The genome sequencing project has generated and will contitute to generate enormous amounts of sequence data. Since the first complete genome sequence of bacterium Haemophilus in fluenzae was published in 1995, the complete genome sequences of 2 eukaryotic and about 22 prokaryotic organisms have detemined. Given this everincreasing amounts of sequence information, new strategies are necessary to efficiently pursue the phase of the geome project- the elucidation of gene expression patterns and gene product function on a whole genome scale. In order to assign functional information to the genome sequence, DNA chip technology was developed to efficienfly identify the differential expression pattern of indepondent biogical samples. DNA chip provides a new tool for genome expreesion analysis that may revolutionize revolutionize many aspects of human kife including mew surg discovery and human disease diagnostics.

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cDNA Sequence and mRNA Expression of a Novel Serine Protease from the Firefly, Pyrocoelia rufa

  • Lee, Kwang-Sik;Kim, Seong-Ryul;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.5 no.1
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    • pp.103-108
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    • 2002
  • We describe here the cDNA sequence and mRNA expression of a novel serine pretense from the firefly, Pyrocoelia rufa. The 771 bp cDNA encodes for 257 amino acid residues. The deduced protein of P. rufa serine pretense gene contains the catalytic triad and six-conserved cysteine residues. Alignment of the deduced protein of P. rufa serine pretense gene showed 47.4% protein sequence identity to known coleopteran insect Rhyzopertha dominica midgut trpsin-like enzyme. Northern blot analysis revealed that the P. rufa serine pretense is specifically expressed in the midgut of P. rufa larvae.

Electrochemical measurement for analysis of DNA sequence (DNA 염기서열 분석을 위한 전기 화학적 측정법)

  • Jo, Seong-Bo;Hong, Jin-Seop;Kim, Yeong-Mi;Park, Jeong-Ho
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.2
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    • pp.92-97
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    • 2002
  • One of the important roles of a DNA chip is the capability of detecting genetic diseases and mutations by analyzing DNA sequence. For a successful electrochemical genotyping, several aspects should be considered including the chemical treatment of electrode surface, DNA immobilization on electrode, hybridization, choice of an intercalator to be selectively bound to double standee DNA, and an equipment for detecting and analyzing the output signal. Au was used as the electrode material, 2-mercaptoethanol was used for linking DNA to Au electrode, and methylene blue was used as an indicator that can be bound to a double stranded DNA selectively. From the analysis of reductive current of this indicator that was bound to a double stranded DNA on an electrode, a normal double stranded DNA was able to be distinguished from a single stranded DNA in just a few seconds. Also, it was found that the peak reduction current of indicator is proportional to the concentration of target DNA to be hybridized with probe DNA. Therefore, it is possible to realize a sim71e and cheats DNA sensor using the electrochemical measurement for genotyping.

Evidence for a Common Molecular Basis for Sequence Recognition of N3-Guanine and N3-Adenine DNA Adducts Involving the Covalent Bonding Reaction of (+)-CC-1065

  • Park, Hyun-Ju
    • Archives of Pharmacal Research
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    • v.25 no.1
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    • pp.11-24
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    • 2002
  • The antitumor antibiotic (+)-CC-1065 can alkylate N3 of guanine in certain sequences. A previous high-field $^1H$ NMR study on the$(+)-CC-1065d[GCGCAATTG*CGC]_2$ adduct ($^*$ indicates the drug alkylation site) showed that drag modification on N3 of guanine results in protonation of the cross-strand cytosine [Park, H-J.; Hurley, L. H. J. Am. Chem. Soc.1997, 119,629]. In this contribution we describe a further analysis of the NMR data sets together with restrained molecular dynamics. This study provides not only a solution structure of the (+)-CC-1065(N3- guanine) DNA duplex adduct but also new insight into the molecular basis for the sequence- specific interaction between (+)-CC-1065 and N3-guanine in the DNA duplex. On the basis of NOESY data, we propose that the narrow minor groove at the 7T8T step and conformational kinks at the junctions of 16C17A and 18A19T are both related to DNA bending in the drugDNA adduct. Analysis of the one-dimensional $^1H$ NMR (in $H_2O$) data and rMD trajectories strongly suggests that hydrogen bonding linkages between the 8-OH group of the (+)-CC-1065 A-sub-unit and the 9G10C phosphate via a water molecule are present. All the phenomena observed here in the (+)-CC-1065(N3-guanine) adduct at 5'$-AATTG^*$are reminiscent of those obtained from the studies on the (+)-CC-1065(N3-adenine) adduct at $5'-AGTTA^*$, suggesting that (+)-CC-1065 takes advantage of the conformational flexibility of the 5'-TPu step to entrap the bent structure required for the covalent bonding reaction. This study reveals a common molecular basis for (+)-CC-1065 alkylation at both $5'-TTG^*$ and $5'-TTA^*$, which involves a trapping out of sequence-dependent DNA conformational flexibility as well as sequence-dependent general acid and general base catalysis by duplex DNA.

Molecular Authentication of Scrophularia herbs by PCR-RFLP Based on rpl-5 Region of Mitochondrial DNA (현삼속 식물의 종판별을 위한 Mitochondrial DNA의 염기서열 및 PCR-RFLP 분석)

  • Lee, Jeong-Hoon;Jo, Ick-Hyun;Lee, Jei-Wan;Park, Chun-Geun;Bang, Kyong-Hwan;Kim, Hong-Sig;Park, Chung-Berm
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.3
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    • pp.173-179
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    • 2010
  • This study describes an efficient approach to the development of DNA markers for use in distinguishing the Scrophularia species that have been used as useful medicinal crops. In order to distinguish Scrophularia species, DNA sequences of rpl-5 region in mitochondrial DNA of Scrophularia species were analysed for detecting sequence variations, and the PCR-RFLP method was applied for developing practicable DNA marker patterns. Several DNA variations were detected by the sequence comparison of rpl-5 region among Scrophularia species. Genetic relationship analysis of Scrophularia species was carried out based on these DNA variations. DNA variations of rpl-5 region were revealed that it was significantly efficient in genetic relationship analysis of Scrophularia species. In addition, Scrophularia species tested in this study were completely discriminated by four polymorphic genotypes by PCR-RFLP combined with Tsp509 I (^AATT) restriction enzyme. Our results suggested that DNA sequence variations of rpl-5 region were sufficiently useful for genetic relationship analysis of Scrophularia species. Polymorphic genotypes by PCR-RFLP using the Tsp509 I enzyme will be useful for discrimination of Scrophularia species as a practicable DNA markers.