• Title/Summary/Keyword: Genome Analysis

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Clonal plant as experimental organisms - DNA mutation rate evaluation in the radiation contaminated area of Fukushima Daiichi NPP accident

  • KANEKO, Shingo
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.25-25
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    • 2018
  • The Fukushima Daiichi Nuclear Power Plant accident in March 2011 caused severe radioactive contamination in the surrounding environment. Since the accident, much attention has been paid to the biological and genetic consequences of organism inhabiting the contaminated area. The effect of radiation exposure on genetic mutation rates is little known, especially for low doses and in situ conditions. Evaluating DNA mutation by low levels of radiation dose is difficult due to the rare mutation event and lack of sequence information before the accident. In this study, correlations with air dose levels and somatic DNA mutation rates were evaluated using Next Generation Sequencer for the clonal plant, Phyllostachys edulis. This bamboo is known to spread an identical clone throughout Japan, and it has the advantage that we can compare genetic mutation rate among identical clone growing different air dose levels. We collected 94 samples of P. edulis from 14 sites with air dose rates from $0.04{\sim}7.80{\mu}Gy/h$. Their clonal identity was confirmed by analysis using 24 microsatellite markers, and then, sequences among samples were compared by MIG sequence. The sequence data were obtained from 2,718 loci. About ~200,000 bp sequence (80 bp X 2,718 loci) were obtained for each sample, and this corresponds to about 0.01% of the genome sequence of P. edulis. In these sequences, 442 loci showed polymorphism patterns including recent origin mutation, old mutation, and sequence errors. The number of mutations per sample ranged from 0 to 13, and did not correlate with air dose levels. This result indicated that DNA mutations have not accumulated in P. edulis living in the air doses levels less than $10{\mu}Gy/h$. Our study also suggests that mutation rates can be assessed by selecting an appropriate experimental approach and analyzing with next generation sequencer.

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Thermotolerant Transgenic Ginseng (Panax ginseng C.A. Meyer) by Introducing Isoprene Synthase Gene through Agrobacterium tumefaciens-mediated Transformation

  • Kim, Ok-Tae;Hyun, Dong-Yun;Bang, Kyong-Hwan;Jung, Su-Jin;Kim, Young-Chang;Shin, Yu-Su;Kim, Dong-Hwi;Kim, Swon-Won;Seong, Nak-Sul;Cha, Seon-Woo;Park, Hee-Woon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.95-99
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    • 2007
  • The cost of conventional cultivation of ginseng (Panax ginseng C.A. Meyer) is very expensive, because shadow condition should be maintained during cultivation periods owing to inherently weak plant for high-temperature. Therefore, application of plant biotechnology may be possible to overcome these difficulties caused by conventional breeding of ginseng. Transgenic plants were produced via Agrobacterium tumefaciens Gv3101, both carrying the binary plasmid pBI121 mLPISO with nptII and Iso (isoprene synthase) gene. Integration of the transgenes into the P. ginseng nuclear genome was confirmed by PCR analysis using nptII primers and Iso primers. RT-PCR result also demonstrated the foreign isoprene synthase gene in three transgenic plant lines (T1, T3, and T5) which was expressed at the transcriptional level. When whole plants of transgenic ginseng were exposed to high temperature at $46^{\circ}C$ for 1 h, a non-transformed plant was wilted from heat shock, whereas a transgenic plant appeared to remain healthy. We suggest that the introduction of exogenous isoprene synthase is considered as alternative methods far generating thermotolerance ginseng.

Factors Affecting Agrobacterium tumefaciens-mediated Transformation of Panax ginseng C.A. Meyer

  • Kim, Ok-Tae;Jung, Su-Jin;Bang, Kyong-Hwan;Kim, Young-Chang;Shin, Yu-Su;Sung, Jung-Sook;Park, Chun-Geon;Seong, Nak-Sul;Cha, Seon-Woo;Park, Hee-Woon
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.2
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    • pp.100-104
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    • 2007
  • A protocol for the production of transgenic Panax ginseng C.A. Meyer was established via Agrobacterium tumefaciens-mediated genetic transformation of direct somatic embryos. A number of conditions related to the co-cultivation were tested with respect to maximizing transformation efficiency. The results showed that pH of the co-cultivation medium (5.7), the bacterial growth phase (optical density; $OD_{600}$ = 0.8), co-cultivation period (3 days), and acetosyringone concentration $(100\;{\mu}M)$ had positive effects on transformation. Selected plantlets were cultured on the medium at an elevated hygromycin level(30 mg/l). Integration of the transgenes into the P. ginseng nuclear genome was confirmed by PCR analysis using hpt primers and by Southern hybridization using hpt-specific probe. The transgenic plantlets were obtained after 3-month cultivation and did not show any detectable variation in morphology or growth characteristics compared to wild-type plants.

Effects of fission yeast ortholog of THOC5 on growth and mRNA export in fission yeast (THOC5의 분열효모 이종상동체가 생장 및 mRNA export에 미치는 영향)

  • Koh, Eun-Jin;Yoon, Jin Ho
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.435-439
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    • 2015
  • THO/TREX complex plays an important role in transcriptional elongation, mRNA processing, nuclear RNA export, and genome stability. A fission yeast, Schizosaccharomyces pombe, SPBC577.04 gene encoding the ortholog of THOC5, a component of THO/TREX complex, was identified and characterized. The S. pombe thoc5 (spthoc5) is not essential for both growth and mRNA export, but deletion of the spthoc5 gene caused growth defect and slight accumulation of $poly(A)^+$ RNA in the nucleus. And the functional spThoc5-GFP protein is localized mainly in the nucleus. Co-immunoprecipitation analysis showed that the Hpr1(THOC1) protein, an evolutionally well-conserved component of THO/TREX complex, interacted with spThoc5 as well as Tho2(THOC2), another subunit of THO complex. These results suggest that S. pombe Thoc5 as a component of THO/TREX complex is also involved in mRNA export from the nucleus.

Plasma Soluble CD30 as a Possible Marker of Adult T-cell Leukemia in HTLV-1 Carriers: a Nested Case-Control Study

  • Takemoto, Shigeki;Iwanaga, Masako;Sagara, Yasuko;Watanabe, Toshiki
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8253-8258
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    • 2016
  • Elevated levels of soluble CD30 (sCD30) are linked with various T-cell neoplasms. However, the relationship between sCD30 levels and the development of adult T-cell leukemia (ATL) in human T-cell leukemia virus type 1 (HTLV-1) carriers remains to be clarified. We here investigated whether plasma sCD30 is associated with risk of ATL in a nested case-control study within a cohort of HTLV-1 carriers. We compared sCD30 levels between 11 cases (i.e., HTLV-1 carriers who later progressed to ATL) and 22 age-, sex- and institution-matched control HTLV-1 carriers (i.e., those with no progression). The sCD30 concentration at baseline was significantly higher in cases than in controls (median 65.8, range 27.2-134.5 U/mL vs. median 22.2, range 8.4-63.1 U/mL, P=0.001). In the univariate logistic regression analysis, a higher sCD30 (${\geq}30.2U/mL$) was significantly associated with ATL development (odds ratio 7.88 and the 95% confidence intervals 1.35-45.8, P = 0.02). Among cases, sCD30 concentration tended to increase at the time of diagnosis of aggressive-type ATL, but the concentration was stable in those developing the smoldering-type. This suggests that sCD30 may serve as a predictive marker for the onset of aggressive-type ATL in HTLV-1 carriers.

Association of Genetic Variants in ARID5B, IKZF1 and CEBPE with Risk of Childhood de novo B-Lineage Acute Lymphoblastic Leukemia in India

  • Bhandari, Prerana;Ahmad, Firoz;Mandava, Swarna;Das, Bibhu Ranjan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3989-3995
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    • 2016
  • Background: Childhood acute lymphoblastic leukemia (ALL) is a heterogeneous genetic disease and its etiology remains poorly understood. Recent genome wide association and replication studies have highlighted specific polymorphisms contributing to childhood ALL predispositions mostly in European populations. It is unclear if these observations generalize to other populations with a lower incidence of ALL. The current case-control study evaluated variants in ARID5B (rs7089424, rs10821936), IKZF1 (rs4132601) and CEBPE (rs2239633) genes, which appear most significantly associated with risk of developing childhood B-lineage ALL. Materials and Methods: Using TaqMan assays, genotyping was conducted for 162 de novo B-lineage ALL cases and 150 unrelated healthy controls in India. Appropriate statistical methods were applied. Results: Genotypic and allelic frequencies differed significantly between cases and controls at IKZF1-rs4132601 (p=0.039, p=0.015) and ARID5B-rs10821936 (p=0.028, p=0.026). Both rs10821936 (p=0.019; OR 0.67; 95% CI=0.47-0.94) and rs4132601 (p=0.018; OR 0.67; 95%CI 0.48-0.94) were associated with reduced disease risk. Moreover, gender-analysis revealed male-specific risk associations for rs10821936 (p=0.041 CT+CC) and rs4132601 (p=0.005 G allele). Further, ARID5B-rs7089424 and CEBPE-rs2239633 showed a trend towards decreased disease risk but without significance (p=0.073; p=0.73). Conclusions: Our findings provide the first evidence that SNPs ARID5B-rs10821936 and IKZF1-rs4132601 are associated with decreased B-lineage ALL susceptibility in Indian children. Understanding the effects of these variants in different ethnic groups is crucial as they may confer different risk of ALL within different populations.

Differentially Expressed Genes by Inhibition of C-terminal Src Kinase by siRNA in Human Vascular Smooth Muscle Cells and Their Association with Blood Pressure

  • Hong, Kyung-Won;Shin, Young-Bin;Kim, Koan-Hoi;Oh, Berm-Seok
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.102-113
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    • 2011
  • C-terminal SRC kinase (CSK) is a ubiquitously expressed, cytosolic enzyme that phosphorylates and inactivates several SRC family protein tyrosine kinases. Recent genomewide association studies have implicated CSK in the regulation of blood pressure. The current study aim is to determine the blood pressure association of the genes regulated by CSK down-regulation. The CSK mRNA expression was downregulated in vascular smooth muscle cells using small interfering RNA (siRNA). CSK mRNA levels fell by 90% in cells that were treated with CSK siRNA; the RNA from these cells was examined by microarray using the Illumina HumanRef-8 v3 platform, which comprises 24,526 reference mRNA probes. On treatment with CSK siRNA, 19 genes were downregulated by more than 2-fold and 13 genes were upregulated by more than 2-fold. Three (CANX, SLC30A7, and HMOX1) of them revealed more than 3 fold differential expression. Interestingly, the HMOX1 SNPs were associated with diastolic blood pressure in the 7551 Koreans using Korea Association REsource data, and the result was supported by the other reports that HMOX1 linked to blood vessel maintenance. Among the remaining 29 differentially expressed genes, seven (SSBP1, CDH2, YWHAE, ME2, PFTK1, G3BP2, and TUFT1) revealed association with both systolic and diastolic blood pressures. The CDH2 gene was linked to blood pressures. Conclusively, we identified 32 differentially expressed genes which were regulated by CSK reduction, and two (HOMX1 and CDH2) of them might influence the blood pressure regulation through CSK pathway.

Genomic Analysis of miR-21-3p and Expression Pattern with Target Gene in Olive Flounder

  • Jo, Ara;Lee, Hee-Eun;Kim, Heui-Soo
    • Genomics & Informatics
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    • v.15 no.3
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    • pp.98-107
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    • 2017
  • MicroRNAs (miRNAs) act as regulators of gene expression by binding to the 3' untranslated region (UTR) of target genes. They perform important biological functions in the various species. Among many miRNAs, miR-21-3p is known to serve vital functions in development and apoptosis in olive flounder. Using genomic and bioinformatic tools, evolutionary conservation of miR-21-3p was examined in various species, and expression pattern was analyzed in olive flounder. Conserved sequences (5'-CAGUCG-3') in numerous species were detected through the stem-loop structure of miR-21-3p. Thus, we analyzed target genes of miR-21-3p. Among them, 3' UTR region of PPIL2 gene indicated the highest binding affinity with miR-21-3p based on the minimum free energy value. The PPIL2 gene showed high expression levels in testis tissue of the olive flounder, whereas miR-21-3p showed rather ubiquitous expression patterns except in testis tissue, indicating that miR-21-3p seems to control the PPIL2 gene expression in a complementary repression manner in various tissues of olive flounder. Taken together, this current study contributes to infer the target gene candidates for the miR-21-3p using bioinformatics tools. Furthermore, our data offers important information on the relationship between miR-21-3p and target gene for further functional study.

Evaluation of Newly Isolated Klebsiella pneumoniae Strains for the Co-Production of 3-hydroxypropionic acid and 1,3-propanediol from Glycerol (새로이 분리된 Klebsiella pneumoniae 균주들의 글리세롤 기반 3-hydroxypropionic acid 및 1,3-propanediol 동시 생산성 평가)

  • Ko, Yeounjoo;Seol, Eunhee;Sekar, Balaji Sundara;Kwon, Seongjin;Lee, Jaehyeon;Park, Sunghoon
    • KSBB Journal
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    • v.31 no.4
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    • pp.246-255
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    • 2016
  • Co-production of 3-hydroxypropionic acid (3-HP) and 1,3-propanediol (1,3-PDO) was suggested as an innovative strategy to overcome several limitations occurring in the single production of 3-HP from glycerol. In this study, two new isolates of Klebsiella pneumoniae, which produce less lipopolysaccharide (LPS) thus considered less pathogenic than K. pneumoniae DSM 2026, were compared and evaluated for their potential for the co-production of 3-HP and 1,3-PDO. The newly isolated strains showed significantly faster sedimentation rate than DSM, which should be beneficial for downstream processing. Analysis of genome sequences of the isolates confirmed the presence of all genes necessary for glycerol assimilation, 1,3-PDO production and biosynthesis of coenzyme $B_{12}$. Co-production yield was highest under anaerobic condition while cell growth was highest under aerobic condition. Both strains showed similarly good performance for the co-production although J2B gave the slightly higher co-production yield of 0.80 mol/mol than GSC021 (0.75 mol/mol). The evaluation of the newly developed strains presented here should be useful in designing similar evaluation experiments for other microorganisms.

Feature Selection for Anomaly Detection Based on Genetic Algorithm (유전 알고리즘 기반의 비정상 행위 탐지를 위한 특징선택)

  • Seo, Jae-Hyun
    • Journal of the Korea Convergence Society
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    • v.9 no.7
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    • pp.1-7
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    • 2018
  • Feature selection, one of data preprocessing techniques, is one of major research areas in many applications dealing with large dataset. It has been used in pattern recognition, machine learning and data mining, and is now widely applied in a variety of fields such as text classification, image retrieval, intrusion detection and genome analysis. The proposed method is based on a genetic algorithm which is one of meta-heuristic algorithms. There are two methods of finding feature subsets: a filter method and a wrapper method. In this study, we use a wrapper method, which evaluates feature subsets using a real classifier, to find an optimal feature subset. The training dataset used in the experiment has a severe class imbalance and it is difficult to improve classification performance for rare classes. After preprocessing the training dataset with SMOTE, we select features and evaluate them with various machine learning algorithms.