• Title/Summary/Keyword: RNA, Ribosomal, 16S

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Eight unrecorded bacterial species isolated from soil and marine sediment in Korea

  • Kim, Minji;Lee, Ki-Eun;Cha, In-Tae;Lee, Byoung-Hee;Park, Soo-Je
    • Journal of Species Research
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    • v.9 no.4
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    • pp.339-345
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    • 2020
  • The Earth contains billions of microbial species, although the vast majority cannot be cultured in laboratories and are thus considered unidentified and uncharacterized. Extremophiles are microorganisms that thrive in extreme conditions, including temperature, salinity, and pH. Extremophilic microorganisms have provided important insights for biological, metabolic, and evolutionary studies. Between 2017 and 2019, as part of a comprehensive investigation to identify bacterial species in Korea, eight bacterial strains were isolated from marine and non-marine environments in Jeju Island. These strains were cultured under extreme salinity or pH conditions. Phylogenetic analysis using 16S ribosomal RNA(rRNA) gene sequencing indicated that all eight strains belonged to the phyla Gammaproteobacteria, Bacilli, and Alphaproteobacteria. Based on their high 16S rRNA gene sequence similarities(>98.7%) and the formation of strong monophyletic clades with their closest related species, all isolated strains were considered as an unrecorded strain, previously unidentified species. Gram stain reaction, culture conditions, colony and cell morphology, biochemical characteristics, isolation source, and National Institute of Biological Resources(NIBR) IDs are described in this article. The characterization of these unrecorded strains provides information on microorganisms living in Korea.

Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.193-204
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    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

Phylogenetic Relationships among Some Bumblebees (Hymenoptera: Apidae) Common in Korea Inferred from Mitochondrial 16S rRNA Sequences

  • Yoon, Hyung-Joo;Lee, Myeong-Lyeol;Kim, Sam-Eun;Lee, Sang-Beom;Kim, Iksoo;Bae, Jin-Sik;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.51-59
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    • 2004
  • Phylogenetic relationships were analyzed among bumblebees using a portion of mitochondrial (mt) 16S ribosomal RNA (16S rRNA). Eight species of true bumblebees and one species of cuckoo bumblebee (Bombini, Apidae), collected from Korea were included in the analysis. Also, one species of true bumblebee imported from several foreign countries for pollination was included. The length of mt 16S rRNA sequence ranged from 496 bp to 508 bp and sequence divergence ranged from 1.4% (7 bp) to 15.49% (77bp). As expected, a high A+T content was observed (78.5% on average). According to the phylogeny tree derived from parsimony and maximum likelihood analysis, a monphyletic Bombus species, excluding a single cuckoo bumblebee, Psithyrus coreanus, was obtained, but the bootstrap estimate at the node supporting the monophyletic group was very weak (40% or 46%), suggesting a very close relationship of the cuckoo bumblebee to the true bumblebee. Within Bombus species belonging to identical subgenera subgeneric specific clustering was formed with high bootstrap values, implying validity of the subgeneric names of each species: Pyrobombus for B. ardens and B. modeatus; Megabombus for B. consobrinus wittenburgi and B. koreanus; and Bombus s. str. for B. ignitus, B. hypocrita sapporoensis, and B. terrestris.

Evidence for Polyphyletic Origin of the Members of the Subsection IV Cyanobacteria as Determined by 16S rRNA Analysis (16S rRNA 분석에 의한 Subsection IV cyanobacteria 균주들의 다계통성 기원의 증거)

  • Shin, Yong Kook;Seo, Pil-Soo
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1202-1206
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    • 2016
  • Unicellular cyanobacterial strains of Subsections I and II and filamentous cyanobacterial strains of Subsection III have been shown to be polyphyletic, heterocystous strains of Subsections IV and V, both of which were previously reported to be monophyletic. In this study, the small subunit ribosomal RNA (16S rRNA) sequences of 13 strains of cyanobacteria - one strain, Oscillatoria nigro-viridis PCC7112, of the Subsection III, 6 strains including genus Anabaena, Nostoc, Tolypothrix, Calothrix and Scytonema of the Subsection IV, and 6 strains including genus Hapalosiphon, Fischerella and Chlorogloeopsis of the Subsection V - were determined. The phylogenetic analysis of cyanobacteria was carried out using the 16S rRNA sequences. The results of the phylogenetic analyses of 16S rRNA sequences, based on Neighbour-joining, maximum-parsimony, and maximum-likelihood methods, indicated that the members of Subsection IV were not monophyletic but polyphyletic. In addition, the phylogenetic results strongly indicated that the genus Scytonema in Subsection IV could be a common ancestor of heterocystous cyanobacteria in Subsection IV and V. Furthermore, the phylogenetic analyses revealed that the genus Anabaena could be phylogenetically diverse and that cyanobacterial strains in Subsection IV might be polyphyletic, whereas those in Subsection V could be monophyletic, as reported before. The results for the genus Anabaena indicate that it should be reclassified.

Characterizations of the Antimicrobial Resistant Determinants in Proteus spp. Isolated from Humans and Chickens in the Chungcheong Province (충청지역의 사람과 닭으로부터 분리된 Proteus속에 속하는 균주에 존재하는 항균제 내성유전자의 유전형 분석)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.4
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    • pp.327-334
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    • 2016
  • Recently, antimicrobial resistance of pathogenic bacteria has been increasing due to excessive use of antimicrobial agents in both humans and livestock. PCR amplification and nucleotide sequence analyses were conducted to investigate16S ribosomal RNA methyltransferase (RMTase) genes and integrons in P. mirabilis strains isolated from clinical specimens and chickens in an area of the Chungcheong providence. In addition, clonality analysis of P. mirabilis strains was performed using a repetitive extragenic palindromic sequence-based PCR (REP-PCR) method. Of the total 38 P. mirabilis isolates, 7 (18.4%) strains were isolated from clinical specimens contained in the RMTase genes and showed resistance to amikacin, tobramycin, and gentamicin. A total of 23 (60.5%) isolates carried class 1 integrons, but no isolates in our study harbored class 2 and class 3 integrons. Class 1 integrons detected in our study harbored genes encoding resistance to aminoglycosides (aadA2, aadA5, aadA7, and aacCA5), ${\beta}$-lactams ($bla_{PSE}$), erythromycin (ereA), lincosamides (linF), and trimethoprim (dfrA12, dfrA17, and dfrA32). We confirmed that the RMTase genes had spread among only the P. mirabilis isolates from clinical specimens, but class 1 integrons had widely disseminated among P. mirabilis isolates from clinical specimens and chickens. In addition, identical REP-PCR banding patterns were evidenced in only P. mirabilis isolates from chickens. Our results suggest the horizontal spreading of P. mirabilis isolates in the chicken farm. To prevent further spreading of antimicrobial resistant genes among P. mirabilis isolates, monitoring and clinical policing will be required.

Phylogenetic Relationship among the Bumblebees (Hymenoptera: Apidae) Common in Korea Inferred from Mitochondrial 16S rRNA Sequences

  • Yoon, Hyung-Joo;Lee, Myeong-Lyeol;Kim, Sam-Eun;Bae, Jin-Sik;Kim, Iksoo;Lee, Sang-Beom;Kim, Keun-Yong;Jin, Byung-Rae;Sohn, Hung-Dae
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.123-125
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    • 2003
  • We provide a molecular study intending to derive an estimate of the relationships within the genus Bombus, 10 species which span 6 subgenera common collected from several regions of Korea and B. terrestris imported from several foreign countries for pollination using a portion of mitochondrial 16S ribosomal RNA. (omitted)

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Isolation of Temperature-sensitive Mutant Escherichia coli YrdC Involved in Universal t6 A tRNA Synthesis (대장균에서 t6 A tRNA의 생합성에 관여하는 필수 단백질 YrdC의 온도 민감형 돌연변이 분리)

  • Hwang, Jihwan
    • Journal of Life Science
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    • v.28 no.2
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    • pp.257-264
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    • 2018
  • The YrdC superfamily is a group of proteins that are highly conserved in almost all organisms sequenced so far. YrdC in Escherichia coli was suggested to be involved in ribosome biogenesis, translation termination, cold adaptation, and threonylcarbamoyl adenosine formation in tRNA. In this study, to unambiguously demonstrate that yrdC is essential in E. coli, we constructed two yrdC mutant strains of E. coli and examined their phenotypes. In the temperature-sensitive yrdC mutant strain, cell growth stopped almost immediately under nonpermissive conditions and it appeared to accumulate 16S ribosomal RNA precursors without significant accumulation of 30S ribosomal subunits. We also cloned yeast and human homologs and demonstrated that they complement the E. coli yrdC-deletion strain. By mutational study, we demonstrated that the concave surface in the middle of the YrdC protein plays an important role in E. coli, yeast, and human versions. By comparison of two yrdC-deletion strains, we also unambiguously demonstrated that yrdC is essential for viability in E. coli and that the functions of its yeast and human homologs overlap with that of E. coli YrdC.

Redescription of two soil ciliates, Anteholosticha bergeri and Bakuella granulifera, from South Korea

  • Chae, Kyu-Seok;Kim, Kang-San;Min, Gi-Sik
    • Journal of Species Research
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    • v.10 no.1
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    • pp.63-71
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    • 2021
  • Anteholosticha bergeri and Bakuella granulifera were isolated from soil samples collected from Muuidong and Songdo-dong, Incheon and confirmed new to South Korea. Including these two newly recorded species, 11 species of Anteholosticha and four species of Bakuella have been recorded in South Korea to date. Anteholosticha bergeri was discriminated from congeners by following characters: cortical granules, 12-16 macronuclei, 5-8 midventral pairs, 2-3 pretransverse cirri, 4-6 transverse cirri, and three dorsal kineties. Bakuella granulifera was identified by cortical granules, 5-11 buccal cirri, 2-5 frontoterminal cirri, 2-5 midventral cirri rows, and 8-12 transverse cirri. The Korean A. bergeri population corresponds to the Austrian population, except for the number of marginal and transverse cirri, and the Korean B. granulifera population corresponds to the Namibian population, except for body size. In addition, small subunit ribosomal RNA(18S rRNA) gene sequences from both species were determined.

Growth Optimization of Delftia sp. for the Odor Control of Organic Waste (유기성 폐기물의 발생 악취 제거를 위한 Delftia sp.의 성장조건 최적화)

  • Kwon, Hyuk-Ku;Jung, Joon-Oh;Chu, Duk-Sung;Lee, Jang-Hoon
    • Journal of Environmental Health Sciences
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    • v.35 no.5
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    • pp.393-401
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    • 2009
  • We isolated and identified a microorganism which was excellent for ammonia oxidation in the biological control of ammonia gas in odor producing materials from organic composting. The isolated strain was tested for growth characteristics and ammonia elimination efficiency under various conditions of temperature, pH, carbon concentration and ammonia concentration. The strain was isolated from a culture broth used in a $NO_2$ producing test with Griess-Ilosvay reagent. The results of 16S rRNA sequence from the isolated strain by using BLANST (Basic Local Alignment Search Tool) and confirming RDP (Ribosomal Database Project II) and ERRD (The European Ribosomal RNA Database) indicate that the strain is related to Delftia sp. UV-Spectrophotometer (Shimadzu, UVmini-1240) was used as a microbial growth test by measuring turbidity on OD660nm and ammonia concentration was measured by Spectrophotometer (HACH, DR-4000). The optimum growth culture conditions of the ammonia oxidizer Delftia sp. were $30^{\circ}C$, pH 7, glucose concentration 1.00% and $(NH_4)_2SO_4$ 0.5 g/l. Ammonia elimination efficiency was over 94% under the same conditions.

Isolation and characterization of acid-resistanct and halophilic bacteria using cultivation technique in Jeju island (배양기법을 활용한 제주도내 내산 및 호염성 미생물의 분리 및 특성 분석)

  • Han, Bit;Kim, Minji;Ryu, Dajung;Lee, Ki-Eun;Lee, Byoung-Hee;Lee, Eun-Young;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.248-257
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    • 2019
  • In this study, we isolated about 70 bacterial strains from terrestrial and marine environments in Jeju island, and finally, total 21 strains were obtained based on the 16S ribosomal RNA gene sequence analysis. These isolated strains were classified into 16 genera of 5 classes and were identified as an unrecorded species in the Republic of Korea. As a result of the substrate utilization and capability for polymer degradation, the physiological phenotypes for acid resistance and halophilic bacteria were observed to be distinct from each other, except for some acid resistance strains. This study might provide basic information on utilization for indigenous microorganisms.