• Title/Summary/Keyword: EST Analysis

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Gene Expression Profiling of Oilseed Rape Embryos Using Microarray Analysis (Microarray 분석을 이용한 유채 종자성숙단계별 유전자 발현 양상)

  • Roh, Kyung Hee;Park, Jong-Sug;Kim, Jong-Bum;Kim, Hyun Uk;Lee, Kyeong-Ryeol;Kim, Sun Hee
    • Journal of Applied Biological Chemistry
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    • v.55 no.4
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    • pp.227-234
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    • 2012
  • We observed that oil began to accumulate at 25 seed days after flowering (DAF) and reached the maximum potential at 35 seed DAF of oilseed rape, and the greatest weight of 100 seeds was obtained at 35 seed DAF. To survey a broad analysis of gene expression in developing embryos of Brassica napus, the Bn 300k microarray have been constructed. The Bn 300k Microarrary was designed from 80,696 unigenes clustered from 543,448 ESTs and 780 cDNA at NCBI. These arrays have been hybridized in a series of experiments with probes derived from seeds and leaf of B. napus. Approximately 8.5% of the 7,000 genes were expressed as ratios 2-fold higher in seed (25 DAF) than leaves and 0.4% at ratios 10. Also we observed that storage and cell differentiation-related genes were highly expressed at 10 DAF, whereas energy-related genes including fatty acid metabolism were increased up depending on seed maturation using Microarray, which was confirmed by reverse transcriptase polymerase chain reaction. These results suggest that B. napus arrays provide a very useful data set of seed-specific expression that can be further analyzed by examination of the promoter regions of these genes and help our understanding of the complex regulatory network in developing seeds.

Environment isotope aided studies on river water and ground water interaction in the Han River basin (동위원소를 이용한 한강유역의 지하수와 지표수의 연관성에 관한 연구)

  • 안종성;김재성
    • Water for future
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    • v.16 no.4
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    • pp.245-252
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    • 1983
  • Recently river water pollution in Korea is given rise to serious problem in aspect of crop production, drinking well, water contamination and etc. Under these urgent situations, it is prime importance to protect water resources from pollutants. An environmental isotope survey of the groundwater form the shallow alluvial and the underlying crystalline rock aquifer of the Han River Basin has been undertaken, Analysis of the data has I) confirmed the hypothesis that the groundwater from the metropolitan area is recharged from the river whereas that form the non-urbanized region of the Basin is replenished by the infiltrating precipitation; ii) shown that crystalline rock aquifers are recharged by the ground water form the overlying alluvium. Old groundwater is a group of wells with tritium values in the range of 0 to 2 TU. These low values indicate that the water sampled was recharged much ealier, at least a few decades, than the other groundwater samples of higher tritium content. The low values in this region may, in fact, reflect the effect of the impermeable clay layers which impede infilteration from the surface. Stable isotope evidence confirmed that a recharge in the karst area occurs at a significantly greater elevation than that to the alluvial aquifer. An analysis of the tritium level collected over an annual cycle suggests that the residence time of groundwater is probably not more than a few months. There does not appear to be any correlation between the trace level of Zn, Mn and Pb in the groundwater and the mechanism of the recharge.

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Survey of Genes Responsive to Long-Term Heat Stress Using a cDNA Microarray Analysis in Mud Loach (Misgurnus mizolepis) Liver (장기 고온 스트레스에 대한 미꾸라지(Misgurnus mizolepis) 간 조직 내 유전자 발현 반응의 cDNA microarray 분석)

  • Cho, Young Sun;Lee, Sang Yoon;Noh, Choong Hwan;Nam, Yoon Kwon;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.18 no.2
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    • pp.65-77
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    • 2006
  • Gene transcripts potentially responsive to the heat stress were surveyed by cDNA microarray analysis in mud loach (Misgurnus mizolepis). Transcriptional profiles of hepatic tissue in the fish exposed to either $23^{\circ}C$ or $32^{\circ}C$ for 4 weeks were compared each other by 3 replicated hybridization assays using 1,124 unigene clones selected from mud loach liver expressed sequence tags (ESTs). A total of 93 clones showed the substantially increased mRNA levels (>2-fold) in $32^{\circ}C$-exposed group when compared in $23^{\circ}C$control group. It includes various enzymes and proteins involved in energy pathway, protease/protein metabolisms, immune/antioxidant functions, cytoskeleton/cell structure, transport and/or signal transduction. Maximum level of increase was up to 15-fold relative to $23^{\circ}C$ treatment. Heat exposure also resulted in the significant decrease (less than 50% relative to $23^{\circ}C$-exposed fish) of the transcriptional activities in 85 genes. Besides the above categories, yolk protein (vitellogenin) and ribosomal proteins were notably down regulated in the fish exposed to heat stress. A number of novel gene transcripts were also detected in both up-regulated and down-regulated groups.

Analysis of Genes Expressed in Mouse Ovaries of Early Developmental Stages (초기발달 단계의 생쥐 난소에서 발현하는 유전자에 관한 연구)

  • Jeon Eun-Hyun;Yoon Se-Jin;Cha Kwang-Yul;Kim Nam-Hyung;Lee Kyung-Ah
    • Development and Reproduction
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    • v.7 no.2
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    • pp.127-136
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    • 2003
  • The present study was conducted to investigate gene expression profile of mouse ovaries during the primordial-primary follicle transition. We isolated total RNA from mouse ovaries at day1(contains only primordial follicles) and day5(contains both primordial and primary follicles) and synthesized cDNA using annealing control primers(ACP, Seegene, Inc., Seoul, Korea). Using 80 different ACPs for PCR, we cloned, sequenced, and analyzed identities of 41 differentially expressed genes(DEGs). According to BLAST analysis, sequences of 33 clones significantly matched database entries, 4 clones were novel, and 4 clones were ESTs. We selected 8 DEGs with interesting functions, Anx11 and Pepp2-Pending highly expressed in day1 ovary, while Apg3/Autlp-like, BPOZ, Ches1, Kcmf1, NHE3, Nid2, Ninj1, SENP3, Suil-rsl, and TIAP/m-survivin highly expressed in days ovary, and confirmed their different expression between day1 ovaries and days ovaries using semi-quantitative RT-PCR. There was no false positive result. Using in situ hybridization, we found that almost all of genes studied were expressed in the oocyte from primordial follicle stage but expression decreased from primary follicle stage. Meanwhile their expression was increased in cuboidal granulosa cells. Different expression of BPOZ and TIAP/m-survivin between primordial and primary follicles was confirmed by using laser capture microdissection followed by real-time PCR BPOZ and TIAP/m-survivin expressed 4.5 and 3.4 fold higher in primary than primordial follicles, respectively. List of genes obtained from the present study will provide insights for the study of mechanism regulating primordial-primary follicle transition.

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Seismic structural demands and inelastic deformation ratios: Sensitivity analysis and simplified models

  • Chikh, Benazouz;Laouami, Nacer;Mebarki, Ahmed;Leblouba, Moussa;Mehani, Youcef;Kibboua, Abderrahmane;Hadid, Mohamed;Benouar, Djillali
    • Earthquakes and Structures
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    • v.13 no.1
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    • pp.59-66
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    • 2017
  • Modern seismic codes rely on performance-based seismic design methodology which requires that the structures withstand inelastic deformation. Many studies have focused on the inelastic deformation ratio evaluation (ratio between the inelastic and elastic maximum lateral displacement demands) for various inelastic spectra. This paper investigates the inelastic response spectra through the ductility demand ${\mu}$, the yield strength reduction factor $R_y$, and the inelastic deformation ratio. They depend on the vibration period T, the post-to-preyield stiffness ratio ${\alpha}$, the peak ground acceleration (PGA), and the normalized yield strength coefficient ${\eta}$ (ratio of yield strength coefficient divided by the PGA). A new inelastic deformation ratio $C_{\eta}$ is defined; it is related to the capacity curve (pushover curve) through the coefficient (${\eta}$) and the ratio (${\alpha}$) that are used as control parameters. A set of 140 real ground motions is selected. The structures are bilinear inelastic single degree of freedom systems (SDOF). The sensitivity of the resulting inelastic deformation ratio mean values is discussed for different levels of normalized yield strength coefficient. The influence of vibration period T, post-to-preyield stiffness ratio ${\alpha}$, normalized yield strength coefficient ${\eta}$, earthquake magnitude, ruptures distance (i.e., to fault rupture) and site conditions is also investigated. A regression analysis leads to simplified expressions of this inelastic deformation ratio. These simplified equations estimate the inelastic deformation ratio for structures, which is a key parameter for design or evaluation. The results show that, for a given level of normalized yield strength coefficient, these inelastic displacement ratios become non sensitive to none of the rupture distance, the earthquake magnitude or the site class. Furthermore, they show that the post-to-preyield stiffness has a negligible effect on the inelastic deformation ratio if the normalized yield strength coefficient is greater than unity.

Evidence of genome duplication revealed by sequence analysis of multi-loci expressed sequence tagesimple sequence repeat bands in Panax ginseng Meyer

  • Kim, Nam-Hoon;Choi, Hong-Il;Kim, Kyung Hee;Jang, Woojong;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.130-135
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    • 2014
  • Background: Panax ginseng, the most famous medicinal herb, has a highly duplicated genome structure. However, the genome duplication of P. ginseng has not been characterized at the sequence level. Multiple band patterns have been consistently observed during the development of DNA markers using unique sequences in P. ginseng. Methods: We compared the sequences of multiple bands derived from unique expressed sequence tagsimple sequence repeat (EST-SSR) markers to investigate the sequence level genome duplication. Results: Reamplification and sequencing of the individual bands revealed that, for each marker, two bands around the expected size were genuine amplicons derived from two paralogous loci. In each case, one of the two bands was polymorphic, showing different allelic forms among nine ginseng cultivars, whereas the other band was usually monomorphic. Sequences derived from the two loci showed a high similarity, including the same primer-binding site, but each locus could be distinguished based on SSR number variations and additional single nucleotide polymorphisms (SNPs) or InDels. A locus-specific marker designed from the SNP site between the paralogous loci produced a single band that also showed clear polymorphism among ginseng cultivars. Conclusion: Our data imply that the recent genome duplication has resulted in two highly similar paralogous regions in the ginseng genome. The two paralogous sequences could be differentiated by large SSR number variations and one or two additional SNPs or InDels in every 100 bp of genic region, which can serve as a reliable identifier for each locus.

Geographic Variation and Interspecific Hybridization between Two Species of the Genus Agkistrodon (Crotalidae) in Korea (한국산 살모사속(뱀과) 2종의 지리적 변이 및 종간 잡종)

  • 백남극;양서영
    • Animal Systematics, Evolution and Diversity
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    • v.5 no.1
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    • pp.77-88
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    • 1989
  • Genetic and morphometric comparison between two species of the genus Agkistrodon were performed to estimate the degree of genic variation and to clarify the taxonomic status of Cheju population, a suspected hybrid form between A. blomhoffii brevicaudus and A u ussuriensis. A sum of 147 specimens representing six populations of A.b. brevicaudus and two populations of A ussuriensis was used in this study. Out of the 21 loci examined, 5 loci (Adh, Ldh-2, Mdh-2, Mpi, Pgi) were monomorphic with identical mobility in both species. O Other Sioci (Aco, Gp, Fum, [po and Xdh in A b. brevicaudus; Est, Got-l, Gp, Mdh-l and 6Pgd in A ussuriensis) were highly poplymorphic. The degree of genic variation of Ab. b brevicaudus and A ussuriensis was A=1.SS, P=42.1%, HD=0.096, HG=O.l1S, and A= 1.4S, P = 31.6%, HD=0.117, HG =0.121, repectively. These values are twofold higher t than those of other reptilian species reported (Selander, 1976) including Korean species of Rhabdophis tigrina and Elaphe dione (Paik and Yang, 1986, 1987). The average values of t the genetic similarities among six populations of A b. brevicaudus and two populations of A ussuriensis were S=0.919 and S=0.962, respectively, whereas the value between species was S = 0.662. Presumed divergent time estimate (Nei, 1975) of these two species was about 1 1.8 million years ago. The analysis of the geographic variations of various morphological c characters was based on Gloyd’s criteria (Gloyd, 1972). Coloration of tongue and tail tip, t the number of ventral scales, and the number of subcaudal scales are good diagnostic c characters to identify these two species. The pattern of cross band is, however, highly variable within and between populations of both species. No hybrid was detected and the s suspected hybrid form of Cheju population falls within the range of A. ussuriensis in the g genetic and morphometric analyses.

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Screening of Edible Mushrooms for the Production of Lovastatin and its HMG-CoA Reductase Inhibitory Activity (Lovastatin을 생산하는 식용버섯 선발과 HMG-CoA reductase 저해 효과)

  • Lee Jae-Won;Lee Soo-Min;Gwak Ki-Seob;Lee Ji-Yoon;Choi In-Gyu
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.83-88
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    • 2006
  • This research was performed to determine the production of lovastatin and its HMG-CoA reductase activity produced by fruit bodies and mycelial liquid cultures of domestic edible mushrooms (8 fungal strains). By deter-mining TLC analysis for the confirmation of the presence of lovastatin, all the extracts from fruit bodies and mycelial liquid culture showed same Rf value (0.46), whick was identical to that of the standard lovastatin. In order to extract lovastatin from fruit body, the mixture of water/acetonitrile/methanol was chosen as the most effective solvent. Extracts from fruit body and mycelial liquid culture of pleurotus ostreatus produced the high-est lovastatin 0.98 mg/g based on dry biomass, and 21.90 mg/L, respectively. In the inhibition rate of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, the highest was obtained in P. ostreatus as 67.8% among fruit bodies, and the rates of mycelial liquid culture extracts from P. ostreatus and Laetiporus sulphureus were 37.2% and 29.1%, respectively. Unusually L. sulphureus showed high inhibition rate with low content of lovastatin due to the contribution of campesterol and gamma-sitosterol with hypocholesterolemic activity as metabolites.

Seismic structural demands and inelastic deformation ratios: a theoretical approach

  • Chikh, Benazouz;Mebarki, Ahmed;Laouami, Nacer;Leblouba, Moussa;Mehani, Youcef;Hadid, Mohamed;Kibboua, Abderrahmane;Benouar, Djilali
    • Earthquakes and Structures
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    • v.12 no.4
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    • pp.397-407
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    • 2017
  • To estimate the structural seismic demand, some methods are based on an equivalent linear system such as the Capacity Spectrum Method, the N2 method and the Equivalent Linearization method. Another category, widely investigated, is based on displacement correction such as the Displacement Coefficient Method and the Coefficient Method. Its basic concept consists in converting the elastic linear displacement of an equivalent Single Degree of Freedom system (SDOF) into a corresponding inelastic displacement. It relies on adequate modifying or reduction coefficient such as the inelastic deformation ratio which is usually developed for systems with known ductility factors ($C_{\mu}$) and ($C_R$) for known yield-strength reduction factor. The present paper proposes a rational approach which estimates this inelastic deformation ratio for SDOF bilinear systems by rigorous nonlinear analysis. It proposes a new inelastic deformation ratio which unifies and combines both $C_{\mu}$ and $C_R$ effects. It is defined by the ratio between the inelastic and elastic maximum lateral displacement demands. Three options are investigated in order to express the inelastic response spectra in terms of: ductility demand, yield strength reduction factor, and inelastic deformation ratio which depends on the period, the post-to-preyield stiffness ratio, the yield strength and the peak ground acceleration. This new inelastic deformation ratio ($C_{\eta}$) is describes the response spectra and is related to the capacity curve (pushover curve): normalized yield strength coefficient (${\eta}$), post-to-preyield stiffness ratio (${\alpha}$), natural period (T), peak ductility factor (${\mu}$), and the yield strength reduction factor ($R_y$). For illustrative purposes, instantaneous ductility demand and yield strength reduction factor for a SDOF system subject to various recorded motions (El-Centro 1940 (N/S), Boumerdes: Algeria 2003). The method accuracy is investigated and compared to classical formulations, for various hysteretic models and values of the normalized yield strength coefficient (${\eta}$), post-to-preyield stiffness ratio (${\alpha}$), and natural period (T). Though the ductility demand and yield strength reduction factor differ greatly for some given T and ${\eta}$ ranges, they remain take close when ${\eta}>1$, whereas they are equal to 1 for periods $T{\geq}1s$.

Molecular cloning and expression pattern of Metallothionein Gene from the left-handed shell, Physa acuta (왼돌아물달팽이 (Physa acuta) 의 Metallothionein 유전자 클로닝 및 발현양상)

  • Jo, Yong-Hun;Baek, Moon-Ki;Kang, Se-Won;Lee, Jae-Bong;Byun, In-Seon;Choi, Sang-Haeng;Chae, Sung-Hwa;Kang, Jung-Ha;Han, Yeon-Soo;Park, Hong-Seog;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.25 no.3
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    • pp.223-230
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    • 2009
  • Metallothioneins (MTs) play a key role in metallic homeostasis and detoxification in most living organisms. In an attempt to study the biological functions and significance of MT in a snail, we cloned and partially characterized the MT gene from the left-handed snail, Physa acutawhich has been regarded as a potential biomonitering species for fresh water. The complete cDNA sequence of PaMT cDNA was identified from the expressed sequence tag (EST) sequencing project of Physa acuta. The coding region of 180 bp gives 60 amino acid residues including the initiation methionine and termination codon. Clustering and phylogenic analysis of PaMT with other MT amino acid sequences show that it has some identities to Helix pomatia (60%), Arianta arbustorum (58%), Perna viridis (49%), Mytilus edulis (49%), Bathymodiolus azoricus (49%), Bathymodiolus azoricus (48%) and Bathymodiolus sp. FD-2002 (48%). Time dependent induction for PaMT from P. acuta exposed with cadmium (50 ppb) indicated that PaMT was induced at 4-8 hr after exposure. It remains to further develop PaMTas a potential biomarker for water contamination in fresh water.

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