• Title/Summary/Keyword: 16S-rRNA

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PCR Specific Primer for the Detection of Vibrio tapetis (Vibrio tapetis의 검출을 위한 PCR specific primer의 제작)

  • Kim, Yeong-Jin;Lee, Sun-Yi;Cho, Hyo-Jin;You, Sun-Nyung;Kim, Cheol-Min;Choi, Yong-Lark;Park, Byoung-Keun;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.162-165
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    • 2007
  • Brown Ring Disease (BRD) is a bacterial disease caused by Vibrio tapetis which affects cultured clam Ruditapes philippinarum and causes heavy economic losses on Atlantic coasts of france, Spain and England. In this study, to evaluate the effective detection of the pathogen, specific primer set based on 16S ribosomal RNA (rRNA) sequences designed for rapid detection of V. tapetis. Polymerase chain reaction (PCR) with this primer set produced the specific band for each V. tapetis. The length of PCR product using designed primer set of Vbts-F and Vbts-R was about 400 bp. Therefore, these primers will be provided with a basic tool for rapid detection of V. tapetis in the various cases such as examination of imported aquatic products, diagnosis of aquatic organisms, and etc.

Characterization of Fe (III)-Reducing Bacteria Isolated from the Sediment of Chunho Reservoir (천호지 저질토에서 분리한 철환원세균의 특성)

  • 안태영;박재홍;이일규;전은형
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.133-138
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    • 2002
  • Microbial Fe (III) reduction is important for the biogeochemical cycle in the sediment of freshwater system. Also, the Fe (III) reducing mechanism make a model of oxidizing organic compounds and reducing toxic heavy metals, such as chrome or uranium. Thirty-seven strains which have Fe (III) reducing activity were isolated from sediments in lake Soyang and Chunho reservoir. The initial concentration of Fe (II) was the highest in sediments of lake Soyang. However, the highest Fe (III) reducing activity was shown in Chunho reservoir. All isolates were tested for Fe (III) reducing activity. Strains C2 and C3, which were isolated from sediments of Chunho reservoir, showed the highest activity. These strains were tested to see if they utilize various electron donors such as glucose, yeast extract, acetate, ethanol and toluene. Significantly, glucose and yeast extract were used as electron donors. Also these strains were conformed to use humid acid and nitrate as electron accepters. The 16S rRNA sequences of strains C2 and C3 were closely related to Aeromonas hydrophila with 95% similarity.

MicroRNA-328 Inhibits Proliferation of Human Melanoma Cells by Targeting TGFB2

  • Li, Jing-Rong;Wang, Jian-Qin;Gong, Qing;Fang, Rui-Hua;Guo, Yun-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1575-1579
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    • 2015
  • Some microRNAs (miRNAs) have been shown to act as oncogenes or tumor suppressor genes in human melanomas. miR-328 is upregulated in blood cells of melanoma patients compared to in healthy controls. This suggests a role for miR-328 in melanoma that warrants investigation. In this study, we demonstrated miR-328 levels to be dramatically decreased in human melanoma cell lines. Moreover, forced expression of miR-328 inhibited proliferation and induced G1-phase arrest of the SK-MEL-1 melanoma cell line. We identified TGFB2 as a direct target gene for miR-328 using a fluorescent reporter assay and western blotting. Levels of TGFB2 were dramatically increased in human melanoma cell lines and were inversely correlated with the miR-328 expression level. Our findings provide new insights into the mechanisms of human melanoma development, indicating that miR-328 has therapeutic potential for this disease.

Analysis of miRNA expression in the trachea of Ri chicken infected with the highly pathogenic avian influenza H5N1 virus

  • Suyeon Kang;Thi Hao Vu;Jubi Heo;Chaeeun Kim;Hyun S. Lillehoj;Yeong Ho Hong
    • Journal of Veterinary Science
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    • v.24 no.5
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    • pp.73.1-73.16
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    • 2023
  • Background: Highly pathogenic avian influenza virus (HPAIV) is considered a global threat to both human health and the poultry industry. MicroRNAs (miRNA) can modulate the immune system by affecting gene expression patterns in HPAIV-infected chickens. Objectives: To gain further insights into the role of miRNAs in immune responses against H5N1 infection, as well as the development of strategies for breeding disease-resistant chickens, we characterized miRNA expression patterns in tracheal tissues from H5N1-infected Ri chickens. Methods: miRNAs expression was analyzed from two H5N1-infected Ri chicken lines using small RNA sequencing. The target genes of differentially expressed (DE) miRNAs were predicted using miRDB. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were then conducted. Furthermore, using quantitative real-time polymerase chain reaction, we validated the expression levels of DE miRNAs (miR-22-3p, miR-146b-3p, miR27b-3p, miR-128-3p, miR-2188-5p, miR-451, miR-205a, miR-203a, miR-21-3p, and miR-200a3p) from all comparisons and their immune-related target genes. Results: A total of 53 miRNAs were significantly expressed in the infection samples of the resistant compared to the susceptible line. Network analyses between the DE miRNAs and target genes revealed that DE miRNAs may regulate the expression of target genes involved in the transforming growth factor-beta, mitogen-activated protein kinase, and Toll-like receptor signaling pathways, all of which are related to influenza A virus progression. Conclusions: Collectively, our results provided novel insights into the miRNA expression patterns of tracheal tissues from H5N1-infected Ri chickens. More importantly, our findings offer insights into the relationship between miRNA and immune-related target genes and the role of miRNA in HPAIV infections in chickens.

Nocardioides tritolerans sp. nov., Isolated from Soil in Bigeum Island, Korea

  • Dastager, Syed G.;Lee, Jae-Chan;Ju, Yoon-Jung;Park, Dong-Jin;Kim, Chang-Jin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1203-1206
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    • 2008
  • A Gram-positive strain designated as MSL-$14^T$ isolated from a soil sample collected from Bigeum Island, Korea, was subjected to polyphasic taxonomy. The isolate was strictly aerobic. Cells were short rods and motile. Optimum growth temperature and pH was 28$^{\circ}C$ and 7.0, respectively. It was characterized chemotaxonomically as having a cell-wall peptidoglycan type based on LL-2,6-diaminopimelic acid and MK-$8(H_4)$ as the predominant menaquinone. The major fatty acids were iso-$C_{16:0}$, $C_{17:1}$ omega8c, and $C_{18:1}$ omega9c. The G+C content was 67.6 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain MSL-$14^T$ is affiliated to the genus Nocardioides and formed a distinct lineage within the genus. MSL-$14^T$ showed highest sequence similarity to Nocardioides aestuarii JCM $12125^T$, having a similarity of 96.5%. Based on the 16S rRNA gene sequence divergence and phenotypic characteristics, it is proposed that strain MSL-$14^T$ should be classified as representing a novel member of the genus Nocardioides, for which we propose the name Nocardioides tritolerans sp. novo The type strain is strain MSL-$14^T$ (=KCTC $19289^T$=DSM $19320^T$).

Monitoring Bacterial Population Dynamics Using Real-Time PCR During the Bioremediation of Crude-Oil-Contaminated Soil

  • Baek, Kyung-Hwa;Yoon, Byung-Dae;Cho, Dae-Hyun;Kim, Byung-Hyuk;Oh, Hee-Mock;Kim, Hee-Sik
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.339-345
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    • 2009
  • We evaluated the activity and abundance of the crude-oil-degrading bacterium Nocardia sp. H17-1 during bioremediation of oil-contaminated soil, using real-time PCR. The total petroleum hydrocarbon(TPH) degradation rate constants(k) of the soils treated with and without H17-1 were $0.103\;d^{-1}$ and $0.028\;d^{-1}$ respectively. The degradation rate constant was 3.6 times higher in the soil with H17-1 than in the soil without H17-1. In order to detect and quantify the Nocardia sp. H17-1 in soil samples, we quantified the genes encoding 16S ribosomal RNA(16S rRNA), alkane monooxygenase(alkB4), and catechol 2,3-dioxygenase(23CAT) with real-time PCR using SYBR green. The amounts of H17-1 16S rRNA and alkB4 detected increased rapidly up to 1,000-folds for the first 10 days, and then continued to increase only slightly or leveled off. However, the abundance of the 23CAT gene detected in H17-1-treated soil, where H17-1 had neither the 23CAT gene for the degradation of aromatic hydrocarbons nor the catechol 2,3-dioxygenase activity, did not differ significantly from that of the untreated soil($\alpha$=0.05,p>0.22). These results indicated that H17-1 is a potential candidate for the bioaugmentation of alkane-contaminated soil. Overall, we evaluated the abundance and metabolic activity of the bioremediation strain H17-1 using real-time PCR, independent of cultivation.

Five Rare Non-Tuberculous Mycobacteria Species Isolated from Clinical Specimens (임상에서 분리된 희귀 비결핵 마이코박테리아 5종)

  • Park, Young-Kil;Lee, Young-Ju;Yu, Hee-Kyung;Jeong, Mi-Young;Ryoo, Sung-Weon;Kim, Chang-Ki;Kim, Hee-Jin
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.5
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    • pp.331-336
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    • 2010
  • Background: Recently, the rate of infections with non-tuberculous mycobacteria (NTM) has been increasing in Korea. Precise identification of NTM is critical to determination of the pathogen and to target treatment of NTM patients. Methods: Sixty-eight unclassified mycobacteria isolates by rpoB PCR-RFLP assay (PRA) collected in 2008 were analyzed by National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) search after sequencing of 16S rRNA, hsp65, rpoB genes. Results: Nineteen strains of 68 isolates were specified as species after sequencing analysis of 3 gene types. We found 3 M. lentifulavum, 5 M. arupense, 4 M. triviale, 4 M. parascrofulaceum, and one M. obuense. One M. tuberculosis and another M. peregrinum were mutated at the Msp I recognition site needed for rpoB PRA. The remaining 49 isolates did not coincide with identical species at the 3 kinds genes. Conclusion: Sequencing analysis of 16S rRNA, hsp65, rpoB was useful for identification of NTM unclassified by rpoB PRA.

A Method for Comparing Multiple Bacterial Community Structures from 16S rDNA Clone Library Sequences

  • Hur, Inae;Chun, Jongsik
    • Journal of Microbiology
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    • v.42 no.1
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    • pp.9-13
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    • 2004
  • Culture-independent approaches, based on 16S rDNA sequences, are extensively used in modern microbial ecology. Sequencing of the clone library generated from environmental DNA has advantages over fingerprint-based methods, such as denaturing gradient gel electrophoresis, as it provides precise identification and quantification of the phylotypes present in samples. However, to date, no method exists for comparing multiple bacterial community structures using clone library sequences. In this study, an automated method to achieve this has been developed, by applying pair wise alignment, hierarchical clustering and principle component analysis. The method has been demonstrated to be successful in comparing samples from various environments. The program, named CommCluster, was written in JAVA, and is now freely available, at http://chunlab.snu.ac.kr/commcluster/.

Rapid and Efficient Detection of 16SrI Group Areca Palm Yellow Leaf Phytoplasma in China by Loop-Mediated Isothermal Amplification

  • Yu, Shao-shuai;Che, Hai-yan;Wang, Sheng-jie;Lin, Cai-li;Lin, Ming-xing;Song, Wei-wei;Tang, Qing-hua;Yan, Wei;Qin, Wei-quan
    • The Plant Pathology Journal
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    • v.36 no.5
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    • pp.459-467
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    • 2020
  • Areca palm yellow leaf (AYL) disease caused by the 16SrI group phytoplasma is a serious threat to the development of the Areca palm industry in China. The 16S rRNA gene sequence was utilized to establish a rapid and efficient detection system efficient for the 16SrI-B subgroup AYL phytoplasma in China by loop-mediated isothermal amplification (LAMP). The results showed that two sets of LAMP detection primers, 16SrDNA-2 and 16SrDNA-3, were efficient for 16SrI-B subgroup AYL phytoplasma in China, with positive results appearing under reaction conditions of 64℃ for 40 min. The lowest detection limit for the two LAMP detection assays was the same at 200 ag/μl, namely approximately 53 copies/μl of the target fragments. Phytoplasma was detected in all AYL disease samples from Baoting, Tunchang, and Wanning counties in Hainan province using the two sets of LAMP primers 16SrDNA-2 and 16SrDNA-3, whereas no phytoplasma was detected in the negative control. The LAMP method established in this study with comparatively high sensitivity and stability, provides reliable results that could be visually detected, making it suitable for application and research in rapid diagnosis of AYL disease, detection of seedlings with the pathogen and breeding of disease-resistant Areca palm varieties.

Isolation and Identification of Microorganism with Potent Fibrinolytic Activity from Korean Traditional Deonjang (재래식 된장으로부터 혈전용해활성을 나타내는 세균의 분리 및 동정)

  • Hyun Kwang-Wook;Lee Jong-Soo;Ham Jung-Hee;Choi Shin-Yang
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.24-28
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    • 2005
  • A bacterial strain D­1 found to have potent fibrinolytic activity was isolated from Korean traditional Doenjang. It was identified as Bacillus sp. based on its 16S rRNA sequence analysis, morphological and physiological characteristics.