Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.
Park, Cho Rong;Kim, Jeong Hee;Yu, Yong Man;Youn, Young Nam
Korean journal of applied entomology
/
v.55
no.3
/
pp.245-256
/
2016
Elytra of Harmonia axyridis exhibit varied color patterns. In the present study, we deciphered the genetic basis for intraspecific diversity of elytra color patterns in H. axyridis, using amplified fragment length polymorphism (AFLP). Twenty-eight AFLP reactions were performed to generate a total of 2,741 bands. Of these, 20 bands were polymorphic for each color pattern. The polymorphic bands showed differences of genetic character among different color patterns of H. axyridis. Among them, ten candidate AFLP markers were color-linked. S1, S2, and S20 markers were detected in Succinea 1 and 2 variants of H. axyridis, whereas S3 and S5 were specifically detected in the Conspicua variant. S15, S18, and S19 were specific to the Succinea 2 variant. Polymerase chain reaction (PCR) products of these ten AFLP markers were sequenced. BLAST analysis of these sequences against the GenBank database revealed their homology to DNA fragments of unknown function. Based on the color-linked AFLP markers, sequence characterized amplified region (SCAR) markers were designed for PCR amplification of genomic DNA. Of the ten AFLP markers, five were successfully converted into SCAR markers, which could discriminate elytra color polymorphism in H. axyridis.
Journal of Korean Tunnelling and Underground Space Association
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v.17
no.4
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pp.499-511
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2015
A room-and-pillar underground structure is characterized by its grid-type array of galleries. As a result, its construction and economical efficiency can be governed by excavation sequence of galleries. Therefore, this study aims to study the optimum excavation scheme of a room-and-pillar underground structure by considering its various design factors such as ground conditions and excavation sequences. Drill-and-blast method is assumed as a excavation method for a room-and-pillar underground structure. In addition, two kinds of excavation patterns corresponding to a concurrent and a sequential excavation patterns are considered in this study. For the assumed conditions, the structural stability and the construction efficiency based on the number of faces and the travel distance of a jumbo drilling machine are analyzed for the two excavation patterns. Even though the two kinds of excavation patterns show almost the same structural stability as each other, the concurrent excavation pattern is relatively preferable to the sequential excavation pattern in terms of the number of faces in operation and travel distance of a drilling jumbo.
Isolation of a gene and determination of its expression pattern are essential in understanding its function. Among the genes localized in 12ql3, stSG3435 EST was chosen to study its expression pattern. The full-length CDNA was cloned by screening of human brain CDNA library and its sequence was determined by serial deletion followed by automated sequencing of the clones with overlapping fragments. The sequence analysis revealed that stSG 3435 CDNA displayed 100% identity to human MYGI and 86% identity to mouse melanocyte proliferation gene-1 (Gamm 1) originally identified from melanocyte, suggesting that MYGI determined by Northern blot analysis revealed the strongest expression in testes with ubiquitous expression in all the tissues tested. In order to investigate the cellular localization of its protein product, the green fluorescence protein gene was fused into the full-length coding sequence of MYGI, Transfection of the fusion construct followed by confocal microscopy resulted in the green fluorescence signal as a punctate state in cytoplasm indication that MYGI was localized in one of the cellular organelles.
Journal of the Korean Institute of Traditional Landscape Architecture
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v.41
no.1
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pp.21-34
/
2023
This study identified the materials and construction methods of 'Old Wall' in 13 villages which were designated as National Registered Cultural Heritage at the time of designation and examined the their structural changes based on field survey. The results are as follows: First, the 'Old Wall' consisted of 10 Soil-Stone Wall and 5 Stone Wall. At the time of designation, Stone Wall, which was built irregularly by dry-construction of natural stones, is similar in shape, but Soil-Stone Wall showed difference by the construction method of making used stones, joints, and faces. Second, the study extracted the changes of 'Old Wall' by repair and examined the changes of construction methods as well as the substitution and addition of materials of structure. The wall-roof was built with cement roof-tile and asbestos slate which have the advantage improve durability and cost-effectiveness. In addition, tile-mouth soil was added to korean traditional roof-tile to prevent rainwater from flowing in. Besides, to improve constructional convenience, the natural stone of the wall-body was replaced with blast stone, float stone and cut stone. Cement block, cement brick and cement mortar were frequently used to repair as well. As Soil-Stone Wall was transformed from irregular pattern-construction to comb pattern-construction and wet-construction was changed to dry-construction, it caused landscape and structural problems. Also, the layer of cement mortar applied to wall-foundation blocked the flow of rainwater that was induced by dry-construction of natural stones. Third, the study regarded that the problem with the repair of 'Old Wall' may occur as it is located in living space, because the owner of the wall could repair for the minor damages without technical knowledge. In addition, it is difficult for repair companies in charge of maintenance of Cultural Heritage to supply local materials, and it is differential construction specifications are not applied.
Seo Eun Young;Cho Moon-June;Lee Jeung Hoon;Lee Young-Sook;Na Myung-Hoon;Lee Woong-Hee;Kim Jun-Sang;Kim Jae-Sung
Radiation Oncology Journal
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v.19
no.4
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pp.389-396
/
2001
Purpose : To detect differentially expressed genes in the patients with uterine cervical cancer during the radiation therapy. Materials and Methods : In patients with biopsy proven uterine cervical cancer, we took tumor tissue just before radiation therapy and at 40 minutes after external irradiation of 1.8 Gy. Total RNAs isolated from non-irradiated and irradiated tumor tissue samples were analyzed using the differential-display reverse transcription-polymerase chain reaction (DDRT-PCR). Complementary DNA (cDNA) fragments corresponding to differentially expressed messenger RNAs(mRNAs) were eluted, and cloned. The differential expression of the corresponding mRNAs was confirmed by reverse northern blot. Differentially expressed cDNA bands were sequenced. Nucleotide sequence data were analyzed in the Gene Bank and EMBL databases via the BLAST network sewer to identify homologies to known genes or cDNA fragments. Expression pattern of down-regulated clone was examined using RT-PCR in S patients undergoing radiotherapy. Results : We identified 18 differentially expressed bands by DDRT-PCR, which were eluted and cloned. There were 10 up-regulated clones and 1 down-regulated clone in reverse northern blot. One cDNA fragment had homology to chemokine receptor CXCR4, four were identified as Human ESTs in the EMBL database in EST clones. Down-regulated CxCa-11 was also down regulated in all patients. Conclusion : Using the DDRT-PCR, we have identified 10 up-regulated and 1 down-regulated clone(s) in the patients with uterine cervical cancer during the radiation therapy. The clinical relevance and the functions of these genes will be further investigated.
Chun, Jae An;Kim, Se Hee;Cho, Kang Hee;Kim, Dae Hyun;Choi, In Myong;Shin, Il Sheob
Journal of Plant Biotechnology
/
v.42
no.1
/
pp.25-33
/
2015
A PyrcpPR-10 gene with differentially expressed was isolated by using the suppression subtractive hybridization assay between '93-3-98' (highly resistant against scab caused by Venturia nashicola) and 'Sweat Skin'(highly susceptible) and analyzed the expression pattern according to organs and cultivars. The full length of PyrcpPR-10 was cloned as 743bp with 480bp's ORP, and was determined to encode a protein of 159 amino acid residues. On analyzing PyrcpPR-10 gene sequence compared with resistant and susceptible cultivars, 'Hwangsilri' (resistant), 'Gamcheonbae' (moderately resistant), 'Wonhwang' (moderately susceptible), 'Niitaka' (highly susceptible), and 'Sweat Skin' (highly susceptible) had identical gene sequence but 'Bartlett' (highly resistant) showed partly different sequences. The deduced amino acid sequence showed 64 ~ 98% homology and had the GXGGXG motif to known amino acid of other plants PR-10 by the BLAST X analysis. Among several organs or tissues, petal was showed highest expression level of PyrcpPR-10 gene followed by leaf, floral axis, bud, and bark. The expression level of PyrcpPR-10 gene was dramatically increased at 24 hr after inoculation in all cultivars and also up-regulated in accordance with resistant degree of cultivars. While resistant cultivars ('Bartlett', '93-3-98', and 'Hwangsilri') induced relatively high expression level of PyrcpPR-10 gene, susceptible cultivars ('Niitaka', and 'Sweat Skin') showed low expression level. PyrcpPR-10 gene is assumed that it is directly connected with defense mechanisms to pear scab.
Seo, Bo Yoon;Jung, Jin Kyo;Park, Ki Jin;Cho, Jum Rae;Lee, Gwan-Seok;Jung, Chung Ryul
Korean journal of applied entomology
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v.53
no.3
/
pp.247-260
/
2014
To identify the species of Trichogramma occurring in the corn fields of Korea as egg parasitoids of Ostrinia furnacalis, we sequenced the full-length of ITS2 nuclear rDNA from 112 parasitoids collected during this study. As a reference to distinguish species, we also retrieved full-length ITS2 sequences of 60 Trichogramma species from the NCBI GenBank database. On the basis of the size and 3'terminal sequence pattern of the ITS2 sequences, the Trichogramma samples collected in this study were divided into three groups (K-1, -2, and -3). Evolutionary distances (d) within and between groups based on ITS2 sequences were estimated to be ${\leq}0.005$ and ${\geq}0.080$, respectively. In the net average distance between groups or species, the d value between K-1 and T. ostriniae, K-2 and T. dendrolimi, and K-3 and T. confusum was the lowest, with values of 0.016, 0.001, and 0.002, respectively. In the phylogenetic tree, K-1 and K-2 were clustered with T. ostriniae and T. dendrolimi, respectively. However, K-3 was clustered with three different species, namely, T. confusum, T. chilonis, and T. bilingensis. NCBI BLAST results revealed that parasitoids belonging to K-1 and K-2 showed 99% identity with T. ostriniae and T. dendrolimi, respectively. Parasitoids in K-3 collected from Hongcheon showed 99-100% identity with T. confusum and T. chilonis, and one parasitoid in K-3 collected from Gochang had 98% identity with T. bilingensis, T. confusum, and T. chilonis. On the basis of these results, we infer that the species of Trichogramma collected in this study are closely related to T. ostriniae (K-1) and T. dendrolimi (K-2). However, it was not possible to distinguish species of K-3 using the ITS2 sequence alone.
The cautious blasting works had been used with emulsion explosion electric M/S delay caps. Drill depth was from 3m to 6m with Crawler Drill ${\phi}70mm$ on the calcalious sand stone (soft -modelate -semi hard Rock). The total numbers of test blast were 88. Scale distance were induced 15.52-60.32. It was applied to propagation Law in blasting vibration as follows. Propagtion Law in Blasting Vibration $V=K(\frac{D}{W^b})^n$ were V : Peak partical velocity(cm/sec) D : Distance between explosion and recording sites(m) W : Maximum charge per delay-period of eight milliseconds or more (kg) K : Ground transmission constant, empirically determind on the Rocks, Explosive and drilling pattern ets. b : Charge exponents n : Reduced exponents where the quantity $\frac{D}{W^b}$ is known as the scale distance. Above equation is worked by the U.S Bureau of Mines to determine peak particle velocity. The propagation Law can be catagorized in three groups. Cubic root Scaling charge per delay Square root Scaling of charge per delay Site-specific Scaling of charge Per delay Plots of peak particle velocity versus distoance were made on log-log coordinates. The data are grouped by test and P.P.V. The linear grouping of the data permits their representation by an equation of the form ; $V=K(\frac{D}{W^{\frac{1}{3}})^{-n}$ The value of K(41 or 124) and n(1.41 or 1.66) were determined for each set of data by the method of least squores. Statistical tests showed that a common slope, n, could be used for all data of a given components. Charge and reduction exponents carried out by multiple regressional analysis. It's divided into under loom over loom distance because the frequency is verified by the distance from blast site. Empirical equation of cautious blasting vibration is as follows. Over 30m ------- under l00m ${\cdots\cdots\cdots}{\;}41(D/sqrt[2]{W})^{-1.41}{\;}{\cdots\cdots\cdots\cdots\cdots}{\;}A$ Over 100m ${\cdots\cdots\cdots\cdots\cdots}{\;}121(D/sqrt[3]{W})^{-1.66}{\;}{\cdots\cdots\cdots\cdots\cdots}{\;}B$ where ; V is peak particle velocity In cm / sec D is distance in m and W, maximLlm charge weight per day in kg K value on the above equation has to be more specified for further understaring about the effect of explosives, Rock strength. And Drilling pattern on the vibration levels, it is necessary to carry out more tests.
ISSR PCR technique was applied to investigate genetic relationship among 5 Mountain cultivated ginseng populations (Jinan, Hongcheon, Punggi, Andong and Yeongju) and cDNA libraries of wild ginseng roots were constructed and analyzed functional genes related to morphogenesis via EST. Twenty four ISSR markers tested produced 127 polymorphic loci from 5 regional Mountain cultivated ginseng. Among the regional samples, Yeongju was made 18 polymorphic loci that were the highest level of variations among the cultivated regions. The range of similarity coefficient was 0.46~0.58 and the regional samples of Punggi and Hongcheon, Jinan and Andong were classified to similar groups respectively, whereas Yeongju was shown to be separate group with high level of genetic variation in UPGMA cluster analysis. As a result, there was no relationship according to geographical distance and genetic similarity. Eleven cDNA clones were consisted of 9 known genes and 2 unknown genes analyzed by BLAST program of NCBI. To recognize expression pattern of Homeodomain transcription factor related genes, Northern Blot analysis was performed for wild ginseng's leaf and root. As a result, the gene was only expressed by Mountain wild ginseng root.
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