• Title/Summary/Keyword: Sequencing Strategy

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Transcriptome Analysis of Antrodia cinnamomea Mycelia from Different Wood Substrates

  • Jiao-Jiao Chen;Zhang Zhang;Yi Wang;Xiao-Long Yuan;Juan Wang;Yu-Ming Yang;Yuan Zheng
    • Mycobiology
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    • v.51 no.1
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    • pp.49-59
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    • 2023
  • Antrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.

Regulation of glucose and glutamine metabolism to overcome cisplatin resistance in intrahepatic cholangiocarcinoma

  • So Mi Yang;Jueun Kim;Ji-Yeon Lee;Jung-Shin Lee;Ji Min Lee
    • BMB Reports
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    • v.56 no.11
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    • pp.600-605
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    • 2023
  • Intrahepatic cholangiocarcinoma (ICC) is a bile duct cancer and a rare malignant tumor with a poor prognosis owing to the lack of an early diagnosis and resistance to conventional chemotherapy. A combination of gemcitabine and cisplatin is the typically attempted first-line treatment approach. However, the underlying mechanism of resistance to chemotherapy is poorly understood. We addressed this by studying dynamics in the human ICC SCK cell line. Here, we report that the regulation of glucose and glutamine metabolism was a key factor in overcoming cisplatin resistance in SCK cells. RNA sequencing analysis revealed a high enrichment cell cycle-related gene set score in cisplatin-resistant SCK (SCK-R) cells compared to parental SCK (SCK WT) cells. Cell cycle progression correlates with increased nutrient requirement and cancer proliferation or metastasis. Commonly, cancer cells are dependent upon glucose and glutamine availability for survival and proliferation. Indeed, we observed the increased expression of GLUT (glucose transporter), ASCT2 (glutamine transporter), and cancer progression markers in SCK-R cells. Thus, we inhibited enhanced metabolic reprogramming in SCK-R cells through nutrient starvation. SCK-R cells were sensitized to cisplatin, especially under glucose starvation. Glutaminase-1 (GLS1), which is a mitochondrial enzyme involved in tumorigenesis and progression in cancer cells, was upregulated in SCK-R cells. Targeting GLS1 with the GLS1 inhibitor CB-839 (telaglenastat) effectively reduced the expression of cancer progression markers. Taken together, our study results suggest that a combination of GLUT inhibition, which mimics glucose starvation, and GLS1 inhibition could be a therapeutic strategy to increase the chemosensitivity of ICC.

Detection of Xanthomonas hortorum pv. carotae in Jeju Island Soils after Carrot Harvest (수확 후 제주 당근 재배 토양에서 Xanthomonas hortorum pv. carotae 분리)

  • Mi-Jin Kim;Hyun Su Kang;Yong Ho Shin;Yong Chull Jeun
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.433-439
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    • 2023
  • Bacterial leaf blight in carrot is one of the most important diseases in the worldwide. In the past decade, its introduction into Korea is causing great concern due to the potential damage to carrot crops domestically. This bacterial disease is caused by Xanthomonas hortorum pv. carotae (Xhc). This study aimed to isolate and identify bacterial strains from the soil of carrot farms in Jeju Island. The bacterial isolates showing characteristics similar with those of Xhc were selected when cultured on artificial media. Through DNA sequencing and analysis based on NCBI data, some of the selected bacterial strains were identified as Xhc. Furthermore, the bacterial strains caused the typical symptom of bacterial leaf blight after inoculation on carrot leaves. The results of this study showed the potential establishment of Xhc in the soil of Jeju Island and it may be valuable data for establish a strategy preventing the domestic spread of carrot bacterial leaf blight in the future.

Metabolic Responses of Activated Sludge to Pentachlorophenol in SBR Systems

  • ;Larry D. Benefield
    • Journal of Environmental Science International
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    • v.3 no.3
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    • pp.273-284
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    • 1994
  • The primary objective of this study was to examine the toxic effects of PCP on activated sludge and to analyze its metabolic responses while treating wastewater containing pentachlorophenol (PCP) in a sequencing batch reactor (SBR) system operating under different control strategies. This study was conducted in two phases 1 and 2 (8-hr and 12-hr cycles). Each phase was operated with two control strategies I and II. Strategy I (reactor 1) involved rapid addition (5 minutes to complete) of substrate to the reactor with continuous mixing but no aeration for 2 hours. Strategy ll (reactor 2) involved adding the feed continuously during the first 2 hours of the cycle when the system was mixed but not aerated. During both phases each reactor was operated at a sludge age of 15 days. The synthetic wastewater was used as a feed. The COD of the feed solution was about 380 mg/l. After the reference response for both reactors was established, the steady state response of each system was established for PCP feed concentrations of 0.1 mg/l, 1.0 mg/l, and 5.0 mg/l in SBR systems operating on both 8-hr and 12-hr cycles. Soluble COD removal was not inhibited at any feed PCP concentrations used. At 5.0 mg/l fined PCP concentration and in SBR systems operating on phase 2, the concentrations of MLVSS were decreased; selective pressure on the mixed biomass might be increased, narrowing the range of possible ecological responses; the settleability of activated sludge was poor; the SOURS were increased, showing that the systems were shocked. Nitrification was made to some extent at all concentrations of feed PCP in SBR systems operating on phase 2 whereas in SBR systems operating on phase 1 little nitrification was observed. Then, nitrification will be delayed as much as soluble COD removal is retarded due to PCP inhibition effects. Enhanced biological phosphorus removal occurring in the system operating with control strategy I during phase 1 of this work and in the presence of low concentrations of PCP was unreliable and might cease at anytime, whereas enhanced biological phosphorus removal occurring in the system operating with either control strategy I or II during phase 2 of this work and in the Presence of feed PCP concentrations up to 1.0 mg/l was reliable. When, however, such processes were exposed to 5.0 mg/l PCP dose, enhanced phosphorus removal ceased and never returned.

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Enterococcus faecium LKE12 Cell-Free Extract Accelerates Host Plant Growth via Gibberellin and Indole-3-Acetic Acid Secretion

  • Lee, Ko-Eun;Radhakrishnan, Ramalingam;Kang, Sang-Mo;You, Young-Hyun;Joo, Gil-Jae;Lee, In-Jung;Ko, Jae-Hwan;Kim, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1467-1475
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    • 2015
  • The use of microbial extracts containing plant hormones is a promising technique to improve crop growth. Little is known about the effect of bacterial cell-free extracts on plant growth promotion. This study, based on phytohormonal analyses, aimed at exploring the potential mechanisms by which Enterococcus faecium LKE12 enhances plant growth in oriental melon. A bacterial strain, LKE12, was isolated from soil, and further identified as E. faecium by 16S rDNA sequencing and phylogenetic analysis. The plant growth-promoting ability of an LKE12 bacterial culture was tested in a gibberellin (GA)-deficient rice dwarf mutant (waito-C) and a normal GA biosynthesis rice cultivar (Hwayongbyeo). E. faecium LKE12 significantly improved the length and biomass of rice shoots in both normal and dwarf cultivars through the secretion of an array of gibberellins (GA1, GA3, GA7, GA8, GA9, GA12, GA19, GA20, GA24, and GA53), as well as indole-3-acetic acid (IAA). To the best of our knowledge, this is the first study indicating that E. faecium can produce GAs. Increases in shoot and root lengths, plant fresh weight, and chlorophyll content promoted by E. faecium LKE12 and its cell-free extract inoculated in oriental melon plants revealed a favorable interaction of E. faecium LKE12 with plants. Higher plant growth rates and nutrient contents of magnesium, calcium, sodium, iron, manganese, silicon, zinc, and nitrogen were found in cell-free extract-treated plants than in control plants. The results of the current study suggest that E. faecium LKE12 promotes plant growth by producing GAs and IAA; interestingly, the exogenous application of its cell-free culture extract can be a potential strategy to accelerate plant growth.

Whole Genome Sequence of a Korean Isolate (strain 51) of Helicobacter pylori

  • Lee Woo Kon;Cho Myung Je;Baik Seung Chul;Song Jae Young;Park Jeong Uck;Kang Hyung Lyun;Youn Hee Shang;Ko Gyung Hyuck;Rhee Kwang Ho
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.180-182
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    • 2002
  • Substantial genomic diversity has been expected among clinical isolates of H. pylori. We have suggested that the two complete H. pylori genomes already sequenced may be insufficient for providing a discriminatory tool for typing clinical isolates as well as an insight into the genomic diversity, which enable to establish strategy for control of H. pylori infection. In this study, we determine the nucleotide sequence of the entire genome of Korean strain 51 and compare it with two reported genomic sequences to suggest validity for extensive genomic sequencing of H. pylori. The genome of H. pylori 51 consists of a circular chromosome with a size of 1,591,297 bp, which is corresponding to $95.4\%\;and\;96.8\%$ of the 26695 and J99 chromosome length, respectively. We predict that there are 1,454 open reading frames (ORFs) in 51, representing $91.4\%\;and\;97.2\%$ of the reported numbers of ORF of 26695 and J99, respectively. In contrast to 26695 and J99 that have 123 and 65 strain-specific genes, respectively, of the 1,454 genes, only 39 genes are unique to 51. Differences in genomic organization between 51 and each foreign strain were greater than between 2 foreign strains in pair wise entire sequence alignments by BLASTN. Particularly, the extent of genomic rearrangement observed between 51 and 26695 is higher than between 51 and J99. Multiple sequence alignment of orthologous genes among 3 strains showed that 51 is genetically closer to 26695 rather than J99. Phylogenetic analysis of nonsynonymous and synonymous mutation indicated J99 has the longest branch length in the unrooted phylogenetic tree, suggesting that J99 has higher mutation rate than the other 2 strains.

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Association Study Between the Polymorphisms of Exostosin-1 Gene and Economic Traits in Hanwoo (한우 Exostosin-1 유전자의 SNP 탐색 및 경제형질 관련성 분석)

  • Kim, Bum-Soo;Kim, Nam-Kuk;Lee, Seung-Hwan;Cho, Yong-Min;Heo, Kang-Nyeong;Park, Eung-Woo;Yang, Boo-Keun;Yoon, Du-Hak
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.7-13
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    • 2011
  • The aim of this study was to identify the polymorphism on exostosin-1 (EXT1) gene and to associate with economic traits in Hanwoo (Korean cattle). We sequenced for detection of single nucleotide polymorphism (SNP) with 24 unrelated individuals and identified four SNPs (T272196A, C272359T, G290964A and A302092G). Relationship between the genotypes of 583 Hanwoo individuals by PCR-RFLP and economic traits were analyzed by general linear model. In EXT1 gene, there were four SNPs associated with economic traits such as eye muscle area breeding value, marbling score breeding value, backfat and thickness breeding value (p<0.05 to p<0.01). In conclusion, this study indicates an important role of EXT1 gene in determining the meat quality or economic characteristics in Hanwoo.

The Infection of Heterosporis anguillarum in Cultured Shortfin Eel (Anguilla bicolor pacifica) (양식동남아산 뱀장어(Anguilla bicolor pacifica)의 Heterosporis anguillarum 감염)

  • Kim, Jin-Do;Do, Jeong-Wan;Choi, Hye-Sung;Jo, Hyae-In;Lee, Nam-Sil;Kim, Young-Dae
    • Korean Journal of Environmental Biology
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    • v.32 no.4
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    • pp.382-388
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    • 2014
  • Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

A TXOP Sharing Scheme for QoS Strategy of IEEE 802.11ac DL MU-MIMO MAC (IEEE 802.11ac DL MU-MIMO MAC의 QoS 정책을 고려한 TXOP 공유 방안)

  • Lee, Ji-Young;Seok, Seung-Joon
    • Journal of Digital Convergence
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    • v.12 no.10
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    • pp.317-327
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    • 2014
  • To improve the efficiency of wireless channel, IEEE 802.11ac uses the DL MU-MIMO MAC scheme through which an AP transmits multiple frames to different mobile nodes simultaneously. IEEE 802.11ac DL MU-MIMO MAC needs a new step, called as TXOP sharing, between legacy IEEE 802.11n DL SU-MIMO's two operations, the obtaining an EDCA TXOP and the transmitting multiple frames for EDCA TXOP. In the TXOP sharing operation, both wireless channel destinations and frames transmitted for its TXOP period should are determined. So this paper deals with the TXOP sharing for improving IEEE 802.11ac MAC performance. However, the EDCA priority based method mentioned by IEEE 802.11ac standard document not fair among the buffers and the frames of buffers, and occurs in high_loss rate and high_delay about specific buffers. In this paper, we propose a new scheme of the TXOP sharing with sequencing p-AC, s-AC in similar properties, and all S-AC. This method provides a differentiated service without damage of EDCA characteristics.

Association of Microsatellite Marker in FABP4 Gene with Marbling Score and Live Weight in Hanwoo

  • Lee, Seung-Hwan;Cho, Yong-Min;Kim, Hyeong-Cheol;Lim, Da-Jeong;Moon, Hee-Joo;Hong, Seong-Koo;Oh, Sung-Jong;Kim, Tae-Hun;Yoon, Du-Hak;Park, Eung-Woo
    • Journal of Animal Science and Technology
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    • v.52 no.6
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    • pp.475-480
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    • 2010
  • The bovine fatty acid binding protein 4 (FABP4) plays an important role to uptake intracellular fatty acid. It has been previously reported as a positional candidate gene for marbling score in livestock. The re-sequencing of FABP4 gene detected a polymorphic AT repeated sequence in intron II of FABP4 gene. Allelic distribution for this microsatellite marker was examined in other cattle breeds. A total of 8 alleles were detected with diverse repeat units (14 to 21 AT repeat) in Hanwoo and 7 breeds. Of the 8 alleles, the predominant alleles were $[AT]_{16}$, $[AT]_{18}$ and $[AT]_{19}$ in the Hanwoo and 7 cattle breeds. The linear mixed model for genotypic effect (3237AT) on carcass traits showed a significant effect on marbling score (MAR P=0.025) and live weight (LWT; P=0.04) in the 583 Hanwoo cattle population. Live weight (LW) was highest in the homozygous $(AT)_{17}$ genotype ($557.5{\pm}6.94$) and lowest in the heterozygous $(AT)_{16/17}$ genotype ($521.7{\pm}7.70$). On the other hand, the homozygous $(AT)_{17}$ genotype ($3.0{\pm}0.15$) has the highest effect on marbling score and the lowest effect was in homozygous (AT)$_{18}$ genotype ($2.2{\pm}0.15$). The marbling score difference between both groups was 0.8 which is around two times higher than SNP genotype effect on marbling score in Limousin $\times$ Wagyu crosses.