• Title/Summary/Keyword: 16S rRNA analysis

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A report of seven unrecorded bacterial species in Korea, isolated from marine sediment

  • Chi Young Hwang;Eui-Sang Cho;Dong-Hyun Jung;Ki-Eun Lee;In-Tae Cha;Won-Jae Chi;Myung-Ji Seo
    • Journal of Species Research
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    • v.12 no.2
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    • pp.158-164
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    • 2023
  • In March 2021, marine sediment from East Sea samples were suspended in a 2% NaCl solution, and serial dilution was performed in fresh marine and Reasoner's 2A agar. Isolated bacterial strains were identified based on 16S rRNA gene sequences, and showed at least 98.7% sequence similarity with previously reported bacterial species. Finally, seven bacterial strains which were validly published but not reported in Korea, were obtained. These isolates were allocated to the orders Bacillales and Flavobacteriales. The three Flavobacteriales strains are classified into the family Flavobacteriaceae. The other four Bacillales belong to the families Bacillaceae and Paenibacillaceae. The seven unrecorded bacterial strains in this study are classified into seven different genera, which are assigned to Mesobacillus, Paenibacillus, Gramella, Gillisia, Arenibacter, Fictibacillus, and Brevibacillus. During the investigation, the possibility of excavation of various unrecorded species in domestic marine sediment was confirmed. Gram-staining, cell morphology, physiological and basic biochemical characteristics, and phylogenetic analysis were performed in this study and provided in the description of each strain.

Morphological characterization and molecular phylogenetic analysis of Dolichospermum hangangense (Nostocales, Cyanobacteria) sp. nov. from Han River, Korea

  • Choi, Hye Jeong;Joo, Jae-Hyoung;Kim, Joo-Hwan;Wang, Pengbin;Ki, Jang-Seu;Han, Myung-Soo
    • ALGAE
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    • v.33 no.2
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    • pp.143-156
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    • 2018
  • Dolichospermum is a filamentous and heterocytous cyanobacterium that is one of the commonly occurring phytoplanktons in the Han River of Korea. Morphological observations led to the identification of D. planctonicum-like filaments in seasonal water samples. In the present study, we successfully isolated these filaments using culture methods, and examined its morphology using light and scanning electron microscopy. The morphology of the D. planctonicumlike species differed from that of typical D. planctonicum; it had thin cylindrical-shaped akinetes, which were narrower towards the ends than at the center. This morphology is firstly described in the genus Dolichospermum. In addition, the akinetes in the filament developed solitarily and were distant from the heterocytes. Phylogenetic analysis of the 16S rRNA sequences showed that our Dolichospermum clustered with D. planctonicum and D. circinale, which have coiled trichome. However, phylogenetic analysis of the gene encoding rivulose-1,5-bisphosphate carboxylase (rbcLX) clearly separated our species from other Dolichospermum, forming a unique clade. Additionally, structures of D. planctonicum and D. hangangense strains were different type in Box-B and V3 region. These results demonstrated that the new Dolichospermum species was unique in morphology and molecular traits. Therefore, we propose this to be a new species belonging to genus Dolichospermum with the name Dolichospermum hangangense sp. nov.

Inhibition of MicroRNA-15a/16 Expression Alleviates Neuropathic Pain Development through Upregulation of G Protein-Coupled Receptor Kinase 2

  • Li, Tao;Wan, Yingchun;Sun, Lijuan;Tao, Shoujun;Chen, Peng;Liu, Caihua;Wang, Ke;Zhou, Changyu;Zhao, Guoqing
    • Biomolecules & Therapeutics
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    • v.27 no.4
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    • pp.414-422
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    • 2019
  • There is accumulating evidence that microRNAs are emerging as pivotal regulators in the development and progression of neuropathic pain. MicroRNA-15a/16 (miR-15a/16) have been reported to play an important role in various diseases and inflammation response processes. However, whether miR-15a/16 participates in the regulation of neuroinflammation and neuropathic pain development remains unknown. In this study, we established a mouse model of neuropathic pain by chronic constriction injury (CCI) of the sciatic nerves. Our results showed that both miR-15a and miR-16 expression was significantly upregulated in the spinal cord of CCI rats. Downregulation of the expression of miR-15a and miR-16 by intrathecal injection of a specific inhibitor significantly attenuated the mechanical allodynia and thermal hyperalgesia of CCI rats. Furthermore, inhibition of miR-15a and miR-16 downregulated the expression of interleukin-$1{\beta}$ and tumor-necrosis factor-${\alpha}$ in the spinal cord of CCI rats. Bioinformatic analysis predicted that G protein-coupled receptor kinase 2 (GRK2), an important regulator in neuropathic pain and inflammation, was a potential target gene of miR-15a and miR-16. Inhibition of miR-15a and miR-16 markedly increased the expression of GRK2 while downregulating the activation of p38 mitogen-activated protein kinase and $NF-{\kappa}B$ in CCI rats. Notably, the silencing of GRK2 significantly reversed the inhibitory effects of miR-15a/16 inhibition in neuropathic pain. In conclusion, our results suggest that inhibition of miR-15a/16 expression alleviates neuropathic pain development by targeting GRK2. These findings provide novel insights into the molecular pathogenesis of neuropathic pain and suggest potential therapeutic targets for preventing neuropathic pain development.

Effect of Obesity and Diabetes on Alzheimer's APP Gene Expression in Mouse Adipose Tissues (비만 및 당뇨가 생쥐 지방조직에서의 Alzheimer's APP 유전자 발현에 미치는 영향)

  • Kim, Jin-Woo;Lee, Yong-Ho
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1012-1018
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    • 2010
  • The aim of this study was to determine whether Alzheimer's amyloid precursor protein (APP) is dysregulated in adipose tissues of C57BL/6 male mice by high-fat diet (HFD) induced obesity, aging, or streptozotocin (STZ)-induced diabetes. APP mRNA expression was examined by quantitative real-time PCR (QPCR) in subcutaneous (SAT) and epididymal adipose tissues (EAT) from mice in 8 different condition groups. By combining conditions of age (16 weeks/26 weeks of age), diet (normal diet (ND)/high-fat diet), and induction of diabetes (non-diabetic/diabetic), 88 mice were divided into 8 different groups. QPCR demonstrated that APP expression in SAT was significantly increased by about two-fold in HFD-induced obese mice compared to both 16 week-old and 26 week-old mice in the ND group (16 weeks p=0.001; 26 weeks p<0.0001), but no changes in EAT was found. Particular effects of aging on APP gene expression were not observed in either adipose tissue depots. Significantly decreased APP expression was found in SAT in STZ-induced diabetic mice fed on ND or HFD at 16 weeks of age (ND p<0.05; HFD p<0.01). Linear regression analysis demonstrated that APP expression levels correlated with body weight in both the non-diabetic group (R=0.657, p<0.0001, n=39) and the diabetic group (R=0.508, p=<0.0001, n=49), but did not correlate with plasma glucose levels, which suggests that decreased APP expression in STZ-induced diabetic mice is most likely due to weight loss rather than hyperglycemia. These data confirm APP dysregulation by weight changes in humans and suggest a possible role linking midlife obesity with the later development of amyloidogenesis in the brain of older patients with Alzheimer's disease.

생물학적 인 제거용 연속회분식 반응기에서의 미생물 분포 조사

  • Jeon, Che-Ok;Park, Jong-Mun
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.77-80
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    • 2000
  • Various analytical methods such as electron microscopy, quinone analysis, and 16S rDNA sequencing studies were used to investigate the microbial communities and to identify the microorganisms responsible for enhanced biological phosphorus removal (EBPR) in an anaerobic/aerobic sequencing batch reactor (SBR) fed with acetate. Electron photomicrographs showed that oval-shaped microorganisms of about $0.7\;{\sim}\;1\;{\mu}m$ in diameter dominated the microbial sludge. These microorganisms contained polyphosphate granules and glycogen inclusions, which suggests that they are a kind of phosphorus accumulating organism. Quinone and 16S rRNA sequence analyses showed that the members of Proteobacteria beta subclass were the most abundant species, which were affiliated with the Rhodocyclus-likes group. Phylogenetic analysis revealed that the two dominating clones of the beta subclass were most distantly related to Propionivibrio dicarboxylicus DSM 5885 and Rhodocyclus tenuis DSM 109 with about 95% and 96% sequence similarity, respectively. Therefore, it was concluded that the oval-shaped organisms related to the Rhodocyclus-likes group are likely to be responsible for biological phosphorus removal in SBR operation supplied with acetate.

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Bacterial Community Analysis and Antibacterial Activity Isolated from Umbraulva japonica (초록갈파래(Umbraulva japonica)에서 분리한 세균의 군집 구조 분석 및 항균 활성)

  • Kim, Ji-Hyun;Park, So-Hyun;Moon, Kyung-Mi;Kim, Dong-Hwi;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.127-134
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    • 2018
  • In this study, 79 bacterial isolates were collected from the surface of marine algae Umbraulva japonica. As a result of analysis of 16s rRNA gene sequence, the 79 isolated bacteria were divided into 4 major groups: [Proteobacteria (74.69%), Actinobacteria (2.53%), Fimicutes (2.53%), and Bacteroidetes (20.25%)] - 7 classes (Actinobacteria, Flavobacteria, Sphingobacteria, Baciili, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria), 12 orders, 17 families and 31 genera. The newly isolated 3 strains could be novel species because of less than 97% similarity in 16s rRNA sequence. Therefore, it is considered that additional experiments should be conducted together with the standard strain. Analysis of 79 bacterial antibacterial activity against human and fish pathogens, such as Edwardsiella tarda, Vibrio harveyi, Streptococcus iniae, Steptococcus parauberis, Escherichia coli, Steptococcus mutans, Listeria monocytogenes and Vibrio vulnificus, was performed by using the supernatant liquid and pellet. As a result, pellet of UJT9, UJT20 and UJR17 showed antibacterial activity against V. vulnificus, UJR17 also showed antibacterial activity against S. parauberis. UJT7 and UJT20, UJR17 have been identified as Bacillus sp. and Pseudomonas sp. and it may be safely assented that it's beneficial to carry out additional experiments for various applications.

Novel Diagnostic Algorithm Using tuf Gene Amplification and Restriction Fragment Length Polymorphism is Promising Tool for Identification of Nontuberculous Mycobacteria

  • Shin, Ji-Hyun;Cho, Eun-Jin;Lee, Jung-Yeon;Yu, Jae-Yon;Kang, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.323-330
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    • 2009
  • Nontuberculous mycobacteria (NTM) are a major cause of opportunistic infections in immunocompromised patients, making the reliable and rapid identification of NTM to the species level very important for the treatment of such patients. Therefore, this study evaluated the usefulness of the novel target genes tuf and tmRNA for the identification of NTM to the species level, using a PCRrestriction fragment length polymorphism analysis (PRA). A total of 44 reference strains and 17 clinical isolates of the genus Mycobacterium were used. The 741 bp or 744 bp tuf genes were amplified, restricted with two restriction enzymes (HaeIII/MboI), and sequenced. The tuf gene-PRA patterns were compared with those for the tmRNA (AvaII), hsp65 (HaeIII/HphI), rpoB (MspI/HaeIII), and 16S rRNA (HaeIII) genes. For the reference strains, the tuf gene-PRA yielded 43 HaeIII patterns, of which 35 (81.4%) showed unique patterns on the species level, whereas the tmRNA, hsp65, rpoB, and 16S rRNA-PRAs only showed 10 (23.3%), 32 (74.4%), 19 (44.2%), and 3 (7%) unique patterns after single digestion, respectively. The tuf gene-PRA produced a clear distinction between closely related NTM species, such as M. abscessus (557-84-58) and M. chelonae (477-84-80-58), and M. kansasii (141-136-80-63-58-54-51) and M. gastri (141-136-117-80-58-51). No difference was observed between the tuf-PRA patterns for the reference strains and clinical isolates. Thus, a diagnostic algorithm using a tuf gene-targeting PRA is a promising tool with more advantages than the previously used hsp65, rpoB, and 16S rRNA genes for the identification of NTM to the species level.

Phylogenetic Analysis and Diversity of Marine Bacteria Isolated from Rhizosphere Soils of Halophyte in Suncheon Bay (순천만에 자생하는 염생식물 근권에서 유래한 해양세균의 계통학적 분석 및 다양성)

  • You, Young-Hyun;Park, Jong Myong;Lee, Myung-Chul;Kim, Jong-Guk
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.65-78
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    • 2015
  • The bacterial diversity of the rhizosphere soil of S. japonica native to Suncheon bay was analyzed. Ninety two strains showing different morphological characteristics were isolated from the soils around the community of S. japonica. Bacterial diversity and distributions were studied by phylogenetic analysis of the partial 16S rDNA sequences. Ninety two strains were partially sequenced and analyzed phylogenetically. These strains were composed of 5 phyla firmicutes (56.5%), gamma-proteobacteria (29.3%), alpha-proteobacteria (5.4%), actinobacteria (5.4%), bacteroidetes (3.3%) and Shannon’s diversity index (H') were different from each of sampling sites (1.675, 1.924 and 2.04). Eleven isolates were presumed to be novel species candidates based on similarity analysis of the 16s rRNA gene sequences. Overall, Firmicutes and gamma-proteobacteria of the rhizosphere soil of S. japonica showed a high diversity.

Trimming conditions for DADA2 analysis in QIIME2 platform

  • Lee, Seo-Young;Yu, Yeuni;Chung, Jin;Na, Hee Sam
    • International Journal of Oral Biology
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    • v.46 no.3
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    • pp.146-153
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    • 2021
  • Accurate identification of microbes facilitates the prediction, prevention, and treatment of human diseases. To increase the accuracy of microbiome data analysis, a long region of the 16S rRNA is commonly sequenced via paired-end sequencing. In paired-end sequencing, a sufficient length of overlapping region is required for effective joining of the reads, and high-quality sequencing reads are needed at the overlapping region. Trimming sequences at the reads distal to a point where sequencing quality drops below a specific threshold enhance the joining process. In this study, we examined the effect of trimming conditions on the number of reads that remained after quality control and chimera removal in the Illumina paired-end reads of the V3-V4 hypervariable region. We also examined the alpha diversity and taxa assigned by each trimming condition. Optimum quality trimming increased the number of good reads and assigned more number of operational taxonomy units. The pre-analysis trimming step has a great influence on further microbiome analysis, and optimized trimming conditions should be applied for Divisive Amplicon Denoising Algorithm 2 analysis in QIIME2 platform.

Isolation and Characterization of a Bacteriophage Preying an Antifungal Bacterium

  • Rahimi-Midani, Aryan;Kim, Kyoung-Ho;Lee, Seon-Woo;Jung, Sang Bong;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.584-588
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    • 2016
  • Several Bacillus species were isolated from rice field soils, and 16S rRNA gene sequence analysis showed that Bacillus cereus was the most abundant. A strain named BC1 showed antifungal activity against Rhizoctonia solani. Bacteriophages infecting strain BC1 were isolated from the same soil sample. The isolated phage PK16 had an icosahedral head of $100{\pm}5nm$ and tail of $200{\pm}5nm$, indicating that it belonged to the family Myoviridae. Analysis of the complete linear dsDNA genome revealed a 158,127-bp genome with G + C content of 39.9% comprising 235 open reading frames as well as 19 tRNA genes (including 1 pseudogene). Blastp analysis showed that the proteins encoded by the PK16 genome had the closest hits to proteins of seven different bacteriophages. A neighbor-joining phylogenetic tree based on the major capsid protein showed a robust clustering of phage PK16 with phage JBP901 and BCP8-2 isolated from Korean fermented food.