• Title/Summary/Keyword: EST Analysis

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A Numerical Model of Inverse Analysis for Estimating the Clogging in the Underground LPG Storage Cavern (지하 LPG 저장공동에서의 Clogging 추정을 위한 역해석 수치모형)

  • 강태섭;한일영
    • Journal of the Korean Society of Groundwater Environment
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    • v.4 no.3
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    • pp.161-167
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    • 1997
  • A numerical model (SK-EST) for estimating hydraulic conductivity using monitoring data of underground LPG storage cavern was developed. The model calculates hydraulic conductivity from matrix equation which is established from the distribution of hydraulic potential. To verify the applicability of this model, an inverse analysis was performed using the monitoring data of pressure cell of an operating underground LPG storage cavern. And also using the water pressure parker test data which were obtained to look over the operation capability of pressure cell, conductivity variation with depth was estimated using the developed numerical model (SK-EST) and was compared with in situ results.

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Organic Solvent-Tolerant Esterase from Sphingomonas glacialis Based on Amino Acid Composition Analysis: Cloning and Characterization of EstSP2

  • Dachuri, VinayKumar;Lee, ChangWoo;Jang, Sei-Heon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1502-1510
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    • 2018
  • Organic solvent-tolerant (OST) enzymes are widely applied in various industries for their activity and stability in organic solvents, for their higher substrate solubility, and for their greater stero-selectivity. However, the criteria for identifying OST enzymes largely remain undefined. In this study, we compared the amino acid composition of 19 OST esterases with that of 19 non OST esterases. OST esterases have increased the ratio of Ala and Arg residues and decreased the ratio of Asn, Ile, Tyr, Lys, and Phe residues. Based on our amino acid composition analysis, we cloned a carboxylesterase (EstSP2) from a psychrophilic bacterium, Sphingomonas glacialis PAMC 26605, and characterized its recombinant protein. EstSP2 is a substrate specific to p-nitrophenyl acetate and hydrolyzed aspirin, with optimal activity at $40^{\circ}C$; at $4^{\circ}C$, the activity is approximately 50% of its maximum. As expected, EstSP2 showed tolerance in up to 40% concentration of polar organic solvents, including dimethyl sulfoxide, methanol, and ethanol. The results of this study suggest that selecting OST esterases based on their amino acid composition could be a novel approach to identifying OST esterases produced from bacterial genomes.

Experimental and numerical analysis of corrosion-induced cover cracking in reinforced concrete sample

  • Richard, Benjamin;Quiertant, Marc;Bouteiller, Veronique;Delaplace, Arnaud;Adelaide, Lucas;Ragueneau, Frederic;Cremona, Christian
    • Computers and Concrete
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    • v.18 no.3
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    • pp.421-439
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    • 2016
  • Corrosion of embedded reinforcing bars is recognized as being the major cause of deterioration of reinforced concrete structures. With regard to maintenance strategies of concrete nuclear structures, the monitoring of cracking remains of primary importance. Recently, authors have developed a post-treatment technique to extract crack features from continuous computations. In this paper, such technique is applied to carry out a numerical analysis of an accelerated corrosion test. Obtained results allow highlighting specific propagation and failure mechanisms that characterize corrosion-induced cracking.

Development of SNP markers for the identification of apple flesh color based on RNA-Seq data (RNA-Seq data를 이용한 사과 과육색 판별 SNP 분자표지 개발)

  • Kim, Se Hee;Park, Seo Jun;Cho, Kang Hee;Lee, Han Chan;Lee, Jung Woo;Choi, In Myung
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.372-378
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    • 2017
  • For comparison of the transcription profiles in apple (Malus domestica L.) cultivars differing in flesh color expression, two cDNA libraries were constructed. Differences in gene expression between red flesh apple cultivar, 'Redfield' and white flesh apple cultivar, 'Granny Smith' were investigated by next-generation sequencing (NGS). Expressed sequence tag (EST) of clones from the red flesh apple cultivar and white flesh apple cultivar were selected for nucleotide sequence determination and homology searches. High resolution melting (HRM) technique measures temperature induced strand separation of short PCR amplicons, and is able to detect variation as small as one base difference between red flesh apple cultivars and white flesh apple cultivars. We applied high resolution melting (HRM) analysis to discover single nucleotide polymorphisms (SNP) based on the predicted SNP information derived from the apple EST database. All 103 pairs of SNPs were discriminated, and the HRM profiles of amplicons were established. Putative SNPs were screened from the apple EST contigs by HRM analysis displayed specific difference between 10 red flesh apple cultivars and 11 white flesh apple cultivars. In this study, we report an efficient method to develop SNP markers from an EST database with HRM analysis in apple. These SNP markers could be useful for apple marker assisted breeding and provide a good reference for relevant research on molecular mechanisms of color variation in apple cultivars.

Evaluation of Extreme Rainfall based on Typhoon using Nonparametric Monte Carlo Simulation and Locally Weighted Polynomial Regression (비매개변수적 모의발생기법과 지역가중다항식을 이용한 태풍의 극치강우량 평가)

  • Oh, Tae-Suk;Moon, Young-Il;Chun, Si-Young;Kwon, Hyun-Han
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.29 no.2B
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    • pp.193-205
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    • 2009
  • Typhoons occurred in the tropical Pacific region, these might be affected the Korea moving toward north. The strong winds and the heavy rains by the typhoons caused a natural disaster in Korea. In the research, the heavy rainfall events based on typhoons were evaluated quantitative through various statistical techniques. First, probability precipitation and typhoon probability precipitation were compared using frequency analysis. Second, EST probability precipitation was calculated by Empirical Simulation Techniques (EST). Third, NL probability precipitation was estimated by coupled Nonparametric monte carlo simulation and Locally weighted polynomial regression. At the analysis results, the typhoons can be effected Gangneung and Mokpo stations more than other stations. Conversely, the typhoons can be effected Seoul and Inchen stations less than other stations. Also, EST and NL probability precipitation were estimated by the long-term simulation using observed data. Consequently, major hydrologic structures and regions where received the big typhoons impact should be review necessary. Also, EST and NL techniques can be used for climate change by the global warming. Because, these techniques used the relationship between the heavy rainfall events and the typhoons characteristics.

Biochemical Characterization of a Novel Thermostable Esterase from the Metagenome of Dokdo Islets Marine Sediment (독도 심해토 메타게놈 유래 신규 내열성 에스테라아제의 생화학적 특성규명)

  • Lee, Chang-Muk;Seo, Sohyeon;Kim, Su-Yeon;Song, Jaeeun;Sim, Joon-Soo;Hahn, Bum-Soo;Kim, Dong-Hern;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.63-70
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    • 2017
  • A functional screen of 60,672 fosmid metagenomic clones amplified from marine sediment obtained from the Dokdo islets in Korea identified the gene EstES1, whose product, EstES1, displayed lipolytic properties on tributyrin-supplemented media. EstES1 is a 576 amino acid protein with a predicted molecular weight of 59.4 kDa including 37 N-terminal leader amino acids. EstES1 exhibited the highest sequence similarity (44%) to a carboxylesterase found in Haliangium ochraceum DSM14365. Phylogenetic analysis indicated that EstES1 belongs to a currently uncharacterized family of lipases. Within the conserved domain, EstES1 retains the catalytic triad that consists of the consensus penta-peptide motif, GESAG. EstES1 demonstrated a broad substrate specificity toward the long acyl group of ethyl esters (C2-C12), and its optimal activity was recorded toward p-Nitrophenyl butyrate (C4) at pH 9.0 and $40^{\circ}C$ (specific activity of 255.4 U/mg). The enzyme remained stable in the ranges of $60-65^{\circ}C$ and pH 9.0-10.5 and in the presence of methanol, ethanol, isopropanol, and dimethyl sulfoxide. Therefore, EstES1 has potential for use in industrial applications involving high temperature, organic solvents, and/or alkaline conditions.

The EST Study of the Peri-implanting Porcine Embryos (Peri-implanting 단계의 돼지배아 EST 연구)

  • Kwak, In-Seok
    • Journal of Life Science
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    • v.19 no.5
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    • pp.587-592
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    • 2009
  • A dramatic morphological change of embryos occurs at peri-implantation. Maternal and embryonic cross-talk during this period, initiated by signals from embryo(s), provides signals for maternal recognition of pregnancy and establishing and maintaining the pregnancy. However, the cellular, biochemical and genetic processes that direct embryo remodeling in mammalian species are not well studied or understood. In order to identify potential genes responsible for morphological change and cross-talk between embryo and uterus, an initial EST analysis was performed. A catalog of expressed genes (Transcriptome) from the d12 peri-implanting porcine embryos was constructed. Six clones were chosen from the initial ESTs for elucidation of their expression patterns during embryogenesis in early pregnancy. A number of these genes demonstrated unique expression profiles in a tissue, cell-type, and temporal fashion, indicating dynamic regulation of embryonic and endometrial gene expressions at different stages of pregnancy. Cross-talk between the embryo and endometrium of the pregnant uterus has provided a suitable micro-environment for the embryo's rapid and dramatic morphological changing process at the peri-implantation stage.

Transferability of EST SSR-Markers from Foxtail Millet to Barnyard Millet (Echinochloa esculenta)

  • Myung Chul Lee;Yu-Mi Choi;Myoung-Jae Shin;Hyemyeong Yoon;Seong-Hoon Kim
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.45-45
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    • 2020
  • A large number of expressed sequence tags (ESTs) in public databases have provided an opportunity for the systematic development of simple sequence repeat (SSR) markers. EST-SSRs derived from conserved coding sequences show considerable cross-species transferability in related species. In the present study, we assessed the utility of foxtail millet EST-SSRs in barnyard millet. A total of 312 EST-SSRs of foxtail millet were tested using 84 Echinochloa crus-galli germplasm accessions; a high rate of transferability (62%) and 46 primer sets (13%) were shown the polymorphism in barnyard millet. The 13% of functional EST-SSRs) was demonstrated between cereals and barnyard millet. SSR marker profile data were scored for the computation of pairwise distances as well as a Neighbor Joining (NJ) tree of all the genotypes. The averaged values of gene diversity (HE) and polymorphism information content (PIC) were 0.213 and 0.179 within populations, respectively. The 84 barnyard millet germplasm accessions were divided into five different groups, which agreed well with their geographical origins. The exotic 12 accessions of India type barnyard millet (E. frumentacea) were all separated form Korean local collection genotype. The present results provide evidence of divergence between cultured and wild type barnyard, as a millet and grass. The polymorphic SSR markers indicated in this study were of great value in analysis of genetic diversity that can be further used for crop improvement through breeding.

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Prediction of Rice Embryo Proteins using EST-Databases

  • Woo, Sun-Hee;Cho, Seung-Woo;Kim, Tae-Seon;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Jong, Seung-Keun
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.1-7
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    • 2008
  • An attempt was made to link rice embryo proteins to DNA sequences and to understand their functions. One hundred of the 700 spots detected on the embryo 2-DE gels were microsequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were unknown in functions as previously reported (Woo et al. 2002). In addition, twenty-four protein spots with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available rice EST databases at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). The chromosomal location of some proteins were also obtained from the rice genetic map provided by Japanese Rice Genome Research Program (http://rgp.dna.affrc.go.jp). The DNA sequence databases including EST have been reported for rice (Oryza sativa L.) now provides whole or partial gene sequence, and recent advances in protein characterization allow the linking proteins to DNA sequences in the functional analysis. This work shows that proteome analysis could be a useful tool strategy to link sequence information and to functional genomics.

Screening and Characterization of an Esterase from a Metagenomic Library

  • KIM JEONG-NYEO;SEO MYUNG-JI;CHO EUN-AH;LEE SANG-JAE;KIM SEONG-BO;CHEIGH CHAN-ICK;PYUN YU-RYANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1067-1072
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    • 2005
  • A metagenomic library was constructed using a fosmid vector, and total genomic DNA was extracted directly from soil at Cisolok (hot spring area, Indonesia). This library was composed of 10,214 clones and screened for lipolytic enzyme on tributyrin agar plates. An esterase gene (estMa) was subcloned and sequenced from a positive lipolytic active clone. Esterase EstMa was encoded by a 954-bp open reading frame and showed low ($11-33\%$) amino acid similarity to known esterases. The amino acid sequence analysis demonstrated that the enzyme is a new member of lipolytic enzyme family VI. The estMa gene encodes a preprotein of 317 amino acids with a predicted molecular mass of 34,799 Da. The purified enzyme exhibited optimal activity at $50^{\circ}C$ and pH 6.5. The $K_m,\;and\;V_{max}$ values of EstMa for the hydrolysis of p-nitrophenyl valerate were $45.3\;{\mu}M$ and 4.45 U/mg, respectively.