• Title/Summary/Keyword: rDNA sequencing

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Genetic Clarification of Auricularia heimuer Strains Bred and Cultivated in Korea Using the ITS and IGS1 rDNA Region Sequences

  • Nitesh Pant;HyeongJin Noh;Won-Ho Lee;Seong Hwan Kim
    • Mycobiology
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    • v.51 no.2
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    • pp.109-113
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    • 2023
  • Auricularia is one of the broadly cultivated edible mushrooms in Korea. Most of the Korean Auricularia strains used for cultivation and breeding are known as A. auricula-judae. Recently, this species has been reported to belong to a species complex. Therefore, this study was carried out to genetically clarify the bred and cultivated Korean A. auricula-judae strains. The internal transcribed spacer (ITS) and IGS1 rDNA region sequences were determined from 10 A. auricula-judae strains by PCR and sequencing. Variation in the nucleotide sequence and sequence length of the two rDNA regions were found among the seven A. auricula-judae strains. A maximum-likelihood (ML) phylogenetic tree based on the ITS sequences clearly placed all the 10 Korean A. auricula-judae strains in the A. heimuer clade of the A. auriculajudae complex. A. heimuer is diverged from A. auricula-judae. An ML phylogenetic tree based on the IGS1 sequences revealed the close relationship between Korean A. heimuer strains to Chinese A. heimuer strains. But each strain could be distinguishable by the IGS1 sequence. Furthermore, progeny strains in the seven Korean strains could be differentiated from their parental strains by the IGS1 sequence based phylogenetic tree. Our results are expected to be used to complement the distinction of domestic Auricularia cultivars.

Isolation of Fungal Deteriogens Inducing Aesthetical Problems and Antifungal Calcite Forming Bacteria from the Tunnel and Their Characteristics (터널에서 미학적 문제를 야기하는 진균 및 항진균 활성을 가진 탄산칼슘 형성세균의 분리와 특성)

  • Park, Jong-Myong;Park, Sung-Jin;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.287-293
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    • 2011
  • The purpose of this study was to isolate and characterize fungal deteriogens, which induce discoloration of the cement tunnel, and calcite forming bacteria (CFBs), which have antifungal activity against fungal deteriogens. Isolation of mold, bacteria and yeast was performed using several solid media and partially identified using internal transcribed spacer (ITS); 5.8S rRNA gene sequencing and 16s rDNA sequencing. A total of 19 microbial strains were identified with the most widely distributed fungal strain being Cladospirum sphaerospermum. In addition, five bacteria derived from the tunnel were identified as CFBs. Amongst the latter, Bacillus aryabhatti KNUC205 exhibited antifungal activity against Cladospirum sphaerospermum KNUC253 and Aspergillus niger KCTC6906 as concentrated filtered supernatants.

Application of DNA Analysis for Identification of Prey Items on Zooplankton: Selective Treatment Method (기수역 요각류 위내용물 유전자 분석: 소화기관 내외부 유전자의 선택적 처리방법)

  • Chae, Yeon-Ji;Oh, Hye-Ji;Kim, Yong-Jae;Chang, Kwang-Hyeon;Jo, Hyunbin
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.247-256
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    • 2021
  • Understanding the selective feeding behavior of zooplankton on phytoplankton is essential for evaluating the nutrient cycle and energy flow in the food web. Although many studies have been conducted regarding the feeding behaviors of zooplankton through gut content analyses, there are limitations in the visual identification of digested contents using a microscope. DNA techniques have been applied to overcome these limitations since they can detect and amplify small amounts of prey DNA remaining in the gut contents. We designed a method to extract prey DNA from the gut contents of the whole body of the copepod specimen and tested the resolution of DNA identification for the prey phytoplankton. The common brackish species, Sinocalanus tenellus, were collected from Saemangeum Reservoir in different sites and seasons, and gut content DNA was extracted using 2.5% bleach treatment for 2 min for removal of potential contamination sources existing in preserved specimens without dissolution of the body. The sequences of the extracted gut contents were confirmed using BLASTn suite based on the NCBI database. The phytoplankton species detected in the gut showed temporal and spatial differences. Although DNA analysis of small copepod gut contents has been suggested as an effective method to examine the dynamics of primary prey sources at the genus or species level, uncertainties such as misidentification and limitations in the detailed information of the composition still exist.

Isolation of Dextran-producing Leuconostoc Zactis from Kimchi

  • Kim, Bong-Joon;Min, Bong-Hee;Kim, Jeongho;Han, Hong-Ui
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.11-16
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    • 2001
  • Tentative identification of Leuconostoc lactis IH23 isolated from kimchi (a fermented vegetable product) has been described previously with 16S rDNA sequencing (Choi, 1., M. Sc. Thesis Inha Univ.1999). This strain produced the slime identified as dextran based on IR, $\^$13/C- and $^1$H-NMR spectroscopic results. Further study proved that the isolate IH23 belongs to a homogeneous genetic group with L. lactis DSM 20202$\^$T/ and L. argentinum DSM 8581$\^$T/. The results showed DNA-DNA homology of 99-100%, 16S rDNA gene sequence similarity (97.7% ), and a phylogenetic relationship although L. argentinum DSM 8581$\^$T/ had lower homology (80-91%). These data indicate that L. argentinum DSM 8581$\^$T/ and the isolate IH23 belong to an identical species with L. lactis DSM 20202$\^$T/at the genetic level, although in carbohydrate fermentation, the isolate IH23 was mast closely related to L. argentinum DSM 8581$\^$T/ and quite different from L. lactis DSM 20202$\^$T/. Here we first report the isolation of consistent phenotypic variation in Leuconostoc lactis. We also emphasize that the nomenclature of subspecies needs to be differentiated into the three strains mentioned above in Leuconostoc lactis.

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Rapid Detection of the pathogenic agent of Bacterial white enteritis of Larval and Juvenile Stages in Olive flounder (Paralichthys olivaceus) (넙치(Paralichthys olivaceus)자치어 장관백탁증(Bacterial white enteritis) 원인균의 신속 검출)

  • Mun, Yeong-Geon;Park, Geun-Tae;Son, Hong-Ju;Lee, Sang-Hyeon;Lee, Jeong-Min;Heo, Mun-Su
    • Journal of fish pathology
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    • v.17 no.3
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    • pp.159-169
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    • 2004
  • Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

Identification of a Regulatory Region within the luxR Structural Gene in a Marine Symbiotic Bacterium, Vibrio fischeri

  • Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.176-182
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    • 1994
  • The light-organ symbiont of pine cone fish, Vibrio fischeri, senses its presence in the host and responds to environmental changes by differentially expressing its symbiosis-related luminescence genes. The V. fischeri luminescence genes are activated by LuxR protein in the presence of an autoinducer. In an effort to elucidate the mechanism of regulation of luxR, a plasmid containing luxR was mutagenized in vitro with hydroxylamine and a luxR mutant plasmid was isolated by its ability to activate luminescence genes cloned in E. coli in the absence of the autoinducer. The specific base change identified by DNA sequencing was only single base transition at +78 from the transcriptional start of luxR. Based on a Western immunoblot analysis, the nucleotide change directed the synthesis of much higher level of LuxR protein without any amino acid substitutions. The results suggest that the region including the +78th base is presumably internal operator required for autorepression of luxR, and the increased cellular level of LuxR results in activation of luminescence genes by autoinducer independent fashion.

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Sequence Analysis of the Internal Transcribed Spacer of Ribosomal DNA in the Genus Rhizopus

  • Park, You-Jung;Min, Byung-Re
    • Mycobiology
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    • v.33 no.2
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    • pp.109-112
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    • 2005
  • The internal transcribed spacer (ITS) regions including the 3'-end of 18S rRNA gene, 5.8S rRNA gene and the 5'-end of the 28S rRNA gene of Rhizopus spp. were amplified by PCR and analyzed by DNASIS program. Length polymorphism of these region ranged from 564 bp in R. oryzae to 789bp in R. stolonifer. The length and sequence of 5.8S was very conserved with $154{\sim}155\;bp$. The sequence of ITS2 was more variable than that of ITS1. The base substitution rates were ranged from 0 to 0.6069 per site, and higher rate was found in R. stolonifer. In general, transition was usually more frequent than transversion. On the basis of sequencing results, four groups were clustered with value of 61.9% similarity; R. oryzae, R. micros pores, R. homothallicus, and R. stolonifer groups.

A riboprinting scheme for identification of unknown Acanthamoeba isolates at species level

  • Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.1
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    • pp.25-31
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    • 2002
  • We describe a riboprinting scheme for identification of unknown Acanthamoeba isolates at the species level. It involved the use of PCR-RFLP of small subunit ribosomal RNA gene (riboprint) of 24 reference strains by 4 kinds of restriction enzymes. Seven strains in morphological group I and III were identified at species level with their unique sizes of PCR product and riboprint type by Rsa 1. Unique RFCP of 17 strains in group II by Dde I. Taq I and Hae III were classified into: (1) four taxa that were identifiable at the species level. (2) a subgroup of 4 taxa and a pair of 2 taxi that were identical with each other. and (3) a species complex of 7 taxa assigned to A. castellanii complex that were closely related. These results were consistent with those obtained by 18s rDNA sequence analysis. This approach provides an alternative to the rDNA sequencing for rapid identification of a new clinical isolate or a large number of environmental isolates of Acanthamoeba.

Isolation of a Marine Bacterium Capable of Biodegrading Poly(butylene succinate)

  • Lee, Sang Jun;Park, Eun Hee;Han, Yun Hee;Kim, Young Ok;Park, Seong Wook
    • Fisheries and Aquatic Sciences
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    • v.16 no.1
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    • pp.41-44
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    • 2013
  • We developed a poly(butylene succinate) (PBS) indicator plate and isolated a marine bacterial colony capable of biodegrading PBS based on the appearance of a clear zone. Growth of the PBS-2 isolate was observed over 4 days of culture at $37^{\circ}C$ in PBS-tryptone basal liquid medium, but not in PBS-deprived control medium. The PBS-2 isolate was named Paenibacillus sp. PBS-2 based on 16S rDNA gene sequencing. The PBS-biodegrading marine bacterium isolated in this study will contribute to the effective management of PBS waste problems in marine environments.

Characterization and Fibrinolytic Activity of Acetobacter sp. FP1 Isolated from Fermented Pine Needle Extract

  • Park, Jae-Young;Yoon, Seo-Hyeon;Kim, Seong-Sim;Lee, Beom-Gi;Cheong, Hyeong-Sook
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.215-219
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    • 2012
  • The strain KCTC 11629BP, isolated from spontaneously fermented pine needle extract (FPE), showed fibrinolysis activity. The isolated strain was analyzed in physiological and biochemical experiments. Based on 16S rDNA sequencing and phylogenic tree analysis, the strain was identified to be a part of the genus Acetobacter, with Acetobacter senegalensis and Acetobacter tropicalis as the closest phylogenetic neighbors. Based on genotypic and phenotypic results, it was proposed that bacterial strain KCTC 11629BP represents a species of the genus Acetobacter. The strain was thusly named Acetobacter sp. FP1. In conclusion, Acetobacter sp. FP1 isolated from FPE possesses fibrinolytic activity.