• Title/Summary/Keyword: ITS sequencing

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A Study on Transcriptome Analysis Using de novo RNA-sequencing to Compare Ginseng Roots Cultivated in Different Environments

  • Yang, Byung Wook
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.5-5
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    • 2018
  • Ginseng (Panax ginseng C.A. Meyer), one of the most widely used medicinal plants in traditional oriental medicine, is used for the treatment of various diseases. It has been classified according to its cultivation environment, such as field cultivated ginseng (FCG) and mountain cultivated ginseng (MCG). However, little is known about differences in gene expression in ginseng roots between field cultivated and mountain cultivated ginseng. In order to investigate the whole transcriptome landscape of ginseng, we employed High-Throughput sequencing technologies using the Illumina HiSeqTM2500 system, and generated a large amount of sequenced transcriptome from ginseng roots. Approximately 77 million and 87 million high-quality reads were produced in the FCG and MCG roots transcriptome analyses, respectively, and we obtained 256,032 assembled unigenes with an average length of 1,171 bp by de novo assembly methods. Functional annotations of the unigenes were performed using sequence similarity comparisons against the following databases: the non-redundant nucleotide database, the InterPro domains database, the Gene Ontology Consortium database, and the Kyoto Encyclopedia of Genes and Genomes pathway database. A total of 4,207 unigenes were assigned to specific metabolic pathways, and all of the known enzymes involved in starch and sucrose metabolism pathways were also identified in the KEGG library. This study indicated that alpha-glucan phosphorylase 1, putative pectinesterase/pectinesterase inhibitor 17, beta-amylase, and alpha-glucan phosphorylase isozyme H might be important factors involved in starch and sucrose metabolism between FCG and MCG in different environments.

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Comparative Transcriptome Analysis of Caryophyllene-Treated Helicobacter pylori

  • Woo, Hyun Jun;Yang, Ji Yeong;Kwon, Hye Jin;Kim, Hyun Woo;Kim, Sa-Hyun;Kim, Jong-Bae
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.440-448
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    • 2021
  • Helicobacter pylori (H. pylori) establishes long-term infections associated with severe gastric diseases such as peptic ulceration and gastric cancer. Exposure to an antibacterial agent can help regulate the expression levels of its pathogenic genes. In this study, we analyzed the transcriptional changes in H. pylori genes induced by β-caryophyllene. We used next-generation sequencing (NGS) to analyze RNA expression changes, and reverse transcription-polymerase chain reaction (RT-PCR) was performed as required to verify the results. The NGS results showed that 30 out of 1,632 genes were expressed differentially by β-caryophyllene treatment. Eleven genes associated with DNA replication, virulence factors, and T4SS components were significantly downregulated. RT-PCR confirmed that treatment reduced the expression levels of 11 genes. RT-PCR showed the reduced expression of 11 genes (dnaE, dnaN, holB, gyrA, cagA, vacA, secA, flgE, virB2, virB4, and virB8) following β-caryophyllene treatment. These results suggest that β-caryophyllene can modulate various H. pylori pathogenic determinants and be a potential therapeutic agent for H. pylori infection.

Comprehensive Expression Analysis of Triterpenoid Biosynthesis Genes Using Pac-Bio Sequencing and rnaSPAdes assembly in Codonopsis lanceolata

  • Ji-Nam Kang;Si Myung Lee;Mi-Hwa Choi;Chang-Kug Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.253-253
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    • 2022
  • Codonopsis lanceolata (C. lanceolata) has been widely used in East Asia as a traditional medicine to treat various diseases such as bronchitis, convulsions, cough, obesity, and hepatitis. C. lanceolata belonging to Campanulaceae contains bioactive compounds such as polyphenols, saponins, and steroids. However, despite the pharmacological significance of C. lanceolata, the genetic information of this plant is limited and there are few studies of its transcriptome. In this study, we constructed a unigene set of C. lanceolata using Pac-Bio sequencing. Furthermore, the reads generated from Pac-bio and Illumina sequencing were mixed and assembled using rnaSPAdes. All genes involved in the triterpenoid pathway, a major bioactive compounds of C. lanceolata, were searched from the two unigene sets and the expression profiles of these genes were analyzed. The results showed that lupeol, beta-amyrin, and dammarenediol synthesis genes were activated in the leaves and roots of C. lanceolata. In particular, the expression of genes related to lupeol synthesis was relatively high, suggesting that the main triterpenoid of C. lanceolata is lupeol. Transcriptome studies related to lupeol synthesis in C. lanceolata have been rarely reported. Lupeol has been reported to have pharmacological effects such as anti-inflammatory, anti-cancer, and anti-bacterial. This study suggests the importance of C. lanceolata as a lupeol producing plant.

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Fetal Lung Interstitial Tumor: A Comprehensive Case Study with an Emphasis on Next-Generation Sequencing

  • Yoo Jin Jung;Seongyeon Jung;Jiwon Koh;Jaemoon Koh;Yoon Kyung Jeon;Sung-Hye Park;Eun Na Kim;Chang Hyun Kang
    • Journal of Chest Surgery
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    • v.57 no.4
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    • pp.408-412
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    • 2024
  • Fetal lung interstitial tumor (FLIT), which is characterized by immature interstitial cells resembling the fetal lung parenchyma of 20 to 24 weeks of gestation, is a rare respiratory neoplasm. This study presents the first reported FLIT in Korea. It also aims to refine the diagnostic method of FLIT and increase the accuracy of prognostic assessment by using next-generation sequencing to check for anaplastic lymphoma receptor tyrosine kinase (anaplastic lymphoma kinase) gene rearrangement. Although the initial prognosis for FLIT has been promising since its first report in 2010, certain pathological features are associated with poorer outcomes. Therefore, achieving an accurate diagnosis of FLIT is crucial for avoiding unnecessary treatments beyond surgical resection.

A Comparison of the Ability of Fungal Internal Transcribed Spacers and D1/D2 Domain Regions to Accurately Identify Candida glabrata Clinical Isolates Using Sequence Analysis

  • Kang, Min-Ji;Choi, Yoon-Sung;Kim, Sunghyun
    • Biomedical Science Letters
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    • v.24 no.4
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    • pp.430-434
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    • 2018
  • Candida glabrata is the second most prevalent causative agent for candidiasis following C. albicans. The opportunistic yeast, C. glabrata, is able to cause the critical bloodstream infections in hospitalized patients. Conventional identification methods for yeasts are often time consuming and labor intensive. Therefore, recent studies on sequence-based identification have been conducted. Recently, sequencing the D1/D2 domain of the large subunit ribosomal RNA gene and the internal transcribed spacers (ITS) 1 and ITS2 regions of the ribosomal DNA has proven useful for DNA-based identification of most species of fungi. In the present study, therefore, fungal ITS and D1/D2 domain regions were targeted and analyzed by DNA sequencing for the accurate identification of C. glabrata clinical isolates. A total of 102 C. glabrata clinical isolates from various clinical samples including bloodstream, catheterized urine, bile and other body fluids were used in the study. The results of the DNA sequence analysis showed that the mean standard deviation of species identity percent score between ITS and D1/D2 domain regions was $97.8%{\pm}2.9$ and $99.7%{\pm}0.46$, respectively. These results revealed that the D1/D2 domain region might be a better target for identifying C. glabrata clinical isolates based on DNA sequences than the ITS1 and ITS2 regions. However, in order to evaluate the usefulness of D1/D2 domain region for species identification of all Candida species, other Candida species such as C. albicans, C. tropicalis, C. dubliniensis, and C. krusei should be verified in further studies additionally.

A Study on the Effect of Bioceramics as Biochemosorption Material in Sequencing Batch Reactor (연속회분식 반응조에서 생화학흡착제로서 바이오세라믹의 영향에 관한 연구)

  • Lee, Seunghwan;Islam, M.S.;Kang, Meea
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.3
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    • pp.367-375
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    • 2006
  • Sequencing Batch Reactor (SBR) is well adopted for community wastewater treatment for its simplicity, performance and various advantageous treatment options. SBR is now drawing attention for its process modification such as coupled with membrane bioreactor, reverse osmosis or applying different media to achieve high removal efficiency. This study focused on the improved efficiency of carbon, nitrogen and phosphorous removal by applying zeolite materials called bioceramics to the SBR. Two laboratory-scale SBR units were operated in the same operating conditions - one with bioceramics called Bioceramic SBR (BCSBR) and the other without bioceramics used as control. Routine monitoring of COD, TP, $NH_3-N$, $NO_3-N$ was performed throughout this study. COD removal was about 80% to 100% and phosphorous removal was about 60% in the process whereas $NH_3-N$ removal efficiency was found to be 99.9% in the BCSBR unit. Addition of bioceramics also improved sludge characteristics such as sludge dewaterability, specific gravity and particle size. BCSBR can withstand high ammonia shock loading leading to the better treatment capacity of high ammonia containing wastewater. The cause of improved removal efficiencies within the biological reactor could be attributed to the biochemosorption mechanisms of bioceramics. Absorption/adsorption or desorption capacity of bioceramics was tested through laboratory experiments.

Whole Genome Sequencing of Two Musa Species Towards Disease Resistance and Fiber Quality Improvement

  • John Ivan Pasquil;Richellen Plaza;Roneil Christian Alonday;Damsel Bangcal;Julianne Villela;Antonio, Lalusin;Maria Genaleen Diaz;Antonio Laurena
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.32-32
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    • 2022
  • Abaca (Musa textilis L. Nee) is a native Musa species from the Philippines known for its natural fiber. Abaca fiber a.k.a. Manila hemp extracted from its pseudostems is considered one of the strongest fibers in the world. This is used for commodities such as ropes, papers, and money bills. Abaca is vulnerable to pests and diseases such as the Abaca Bunchy Top Disease (ABTD) caused by Abaca Bunchy Top Virus (ABTV) and Banana Bunchy Top Virus (BBTV). Inosa, one of the varieties of abaca utilized in the Philippines, is highly susceptible to ABTD. In contrast, Pacol (Musa balbisiana L.), a close relative of abaca, is highly resistant to the same disease. Here, we report the sequencing and de novo genome assembly of both abaca var. Inosa and banana var. Pacol. A total of ~16 Gb and ~21 Gb raw reads for Inosa and Pacol, respectively, were generated using Pacbio Hifi sequencing method and assembled with Hifiasm. High-quality de novo assemblies of both Musa species with 99% recovered as per BUSCO analysis were obtained. The assembled Inosa genome has a total length of ~654 Mb and N50 of 7 Mb while Pacol has a total length of 527 Mb and N50 of 3 Mb which are close to their estimated genome size of ~638 Mb and ~503 Mb, respectively. The information that can be derived from the de novo assembled genomes would provide a solid foundation for further research in disease resistance and fiber quality improvement in abaca.

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Whole genome sequence of Staphylococcus aureus strain RMI-014804 isolated from pulmonary patient sputum via next-generation sequencing technology

  • Ayesha, Wisal;Asad Ullah;Waheed Anwar;Carlos M. Morel;Syed Shah Hassan
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.34.1-34.10
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    • 2023
  • Nosocomial infections, commonly referred to as healthcare-associated infections, are illnesses that patients get while hospitalized and are typically either not yet manifest or may develop. One of the most prevalent nosocomial diseases in hospitalized patients is pneumonia, among the leading causes of mortality and morbidity. Viral, bacterial, and fungal pathogens cause pneumonia. More severe introductions commonly included Staphylococcus aureus, which is at the top of bacterial infections, per World Health Organization reports. The staphylococci, S. aureus, strain RMI-014804, mesophile, on-sporulating, and non-motile bacterium, was isolated from the sputum of a pulmonary patient in Pakistan. Many characteristics of S. aureus strain RMI-014804 have been revealed in this paper, with complete genome sequence and annotation. Our findings indicate that the genome is a single circular 2.82 Mbp long genome with 1,962 protein-coding genes, 15 rRNA, 49 tRNA, 62 pseudogenes, and a GC content of 28.76%. As a result of this genome sequencing analysis, researchers will fully understand the genetic and molecular basis of the virulence of the S. aureus bacteria, which could help prevent the spread of nosocomial infections like pneumonia. Genome analysis of this strain was necessary to identify the specific genes and molecular mechanisms that contribute to its pathogenicity, antibiotic resistance, and genetic diversity, allowing for a more in-depth investigation of its pathogenesis to develop new treatments and preventive measures against infections caused by this bacterium.

Inference of Sequencing Batch Reactor Process using Oxidation Reduction Potential (ORP profile을 이용한 연속 회분식 반응기(Sequencing Batch Reactor)에서 무산소공정 추론)

  • Sim, Mun Yong;Bu, Gyeong Min;Im, Jeong Hun;U, Hye Jin;Kim, Chang Won
    • Journal of Environmental Science International
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    • v.13 no.3
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    • pp.245-250
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    • 2004
  • The SBR(Sequencing Batch Reactor) process is ideally suited to treat high loading wastewater due to its high dilution rate. SBR operates by a cycle of periods consisting of filling, reacting, settling, decanting and idling. The react phases such as aeration or non-aeration, organic oxidation, nitrification, denitrification and other bio-logical reactions can be achieved in a reactor. Although the whole reactions can be achieved in a SBR with time distributing, it is hard to manage the SBR as a normal condition without recognizing a present state. The present state can be observed with nutrient sensors such as ${NH_{4}}^{+}-N$, ${NO_{2}}^{-}-N$, ${NO_{3}}^{-}-N} and ${PO_{4}}^{ 3-}-P.$ However, there is still a disadvantage to use the nutrient sensors because of their high expense and inconvenience to manage. Therefore, it is very useful to use common on-line sensors such as DO, ORP and pH, which are less expensive and more convient. Moreover, the present states and unexpected changes of SBR might be predicted by using of them. This study was conducted to get basic materials for making an inference of SBR process from ORP(oxidation reduction potential) of synthetic wastewater. The profiles of ORP, DO, and pH were under normal nitrification and denitrification were obtained to compare abnormal condition. And also, nitrite and nitrate accumulation were investigated during reaction of SBR. The bending point on ORP profile was not entirely in the low COD/NOx ratio condition. In this case, NOx was not entirely removed, and minimum ORP value was presented over -300mV. Under suitable COD/NOx ratio which complete denitrification was achieved, ORP bending point was observed and minimum ORP value was under -300m V. Under high COD/NOx ratio, ORP bending point was not detected at the first subcycle because of the fast denitrification and minimum ORP value was under -300mV at the time.

Transcriptome sequencing revealed the inhibitory mechanism of ketoconazole on clinical Microsporum canis

  • Wang, Mingyang;Zhao, Yan;Cao, Lingfang;Luo, Silong;Ni, Binyan;Zhang, Yi;Chen, Zeliang
    • Journal of Veterinary Science
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    • v.22 no.1
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    • pp.4.1-4.13
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    • 2021
  • Background: Microsporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans. Objectives: In clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood. The antifungal mechanism of KTZ needs to be studied in detail. Methods: In this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis. The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto. Results: At half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated (p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ. Conclusions: The results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.