• Title/Summary/Keyword: entire sequence

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Detailed Analysis of the 5'-Coding Region of SCN5A Gene in Korean Genome

  • Yeo, Shin-Il;Kim, Su-Won;Kim, Yoon-Nyun;You, Kwan-Hee;Shin, Song-Woo;Kim, Myoung-Hee;Song, Jae-Chan;Yoo, Min
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.189-193
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    • 2002
  • We have identified and analyzed the 5'-coding region of SCN5A gene in Korean genome. Although its sequence has already been reported in western countries it is still important to confirm our own sequence for the establishment of Korean-suitable diagnosis on genetic basis. Total RNAs were obtained from three healthy Korean adult hearts and reversely transcribed. RT products were then subjected to PCR reaction followed by DNA sequencing. Three different sets of SCN5A primers were designed and used for the amplification of 5'-coding region of SCN5A from Korean genome. Amplified sequence was roughly one-10th of the entire SCN5A mRNA in size and its detailed sequence was completely matched up to the previously reported sequence. There was no difference between three heart samples, either. So, SCN5A was concluded as the relatively stable gene comparing to other genes that are involved in long QT syndrome.

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Safety analysis of marine nuclear reactor in severe accident with dynamic fault trees based on cut sequence method

  • Fang Zhao ;Shuliang Zou ;Shoulong Xu ;Junlong Wang;Tao Xu;Dewen Tang
    • Nuclear Engineering and Technology
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    • v.54 no.12
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    • pp.4560-4570
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    • 2022
  • Dynamic fault tree (DFT) and its related research methods have received extensive attention in safety analysis and reliability engineering. DFT can perform reliability modelling for systems with sequential correlation, resource sharing, and cold and hot spare parts. A technical modelling method of DFT is proposed for modelling ship collision accidents and loss-of-coolant accidents (LOCAs). Qualitative and quantitative analyses of DFT were carried out using the cutting sequence (CS)/extended cutting sequence (ECS) method. The results show nine types of dynamic fault failure modes in ship collision accidents, describing the fault propagation process of a dynamic system and reflect the dynamic changes of the entire accident system. The probability of a ship collision accident is 2.378 × 10-9 by using CS. This failure mode cannot be expressed by a combination of basic events within the same event frame after an LOCA occurs in a marine nuclear reactor because the system contains warm spare parts. Therefore, the probability of losing reactor control was calculated as 8.125 × 10-6 using the ECS. Compared with CS, ECS is more efficient considering expression and processing capabilities, and has a significant advantage considering cost.

Complete Nucleotide Sequence of KCNE1 in Korean Genome

  • Yeo, Shin-Il;Kim, Su-Won;Kim, Yoon-Nyun;You, Kwan-Hee;Shin, Song-Woo;Kim, Myoung-Hee;Song, Jae-Chan;Yoo, Min
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.185-188
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    • 2002
  • We have cloned the gene fur long QT syndrome in Korean genome and determined its detailed nucleotide sequence. Blood DNAs were isolated from 68 healthy individuals (including males and females) and the genomic DNAs were amplified by PCR method followed by automatic DNA sequencing. Entire sequence of the coding region for KCNEI was located in exon 3. PCR products were reexamined for the confirmation of KCNE1-specific amplification by nested PCR. KCNE1 mRNA was 436 bp. This corresponded to 129 amino acids. There was no recognizable difference between males and females. This study should contribute to the better understanding of long QT syndrome in Korean population.

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Isolation of the Inositol Phosphoceramide Synthase Gene (AUR1) from Stress-Tolerant Yeast Pichia kudriavzevii

  • Yoo, Boung-Hyuk;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1902-1907
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    • 2015
  • This study is the first report of the entire nucleotide sequence of an inositol phosphoceramide synthase gene from the stress-tolerant yeast Pichia kudriavzevii (PkAUR1). Sequence analysis revealed an open reading frame that spans 1,443 bp and encodes a 480-amino-acid-residue protein with the highest sequence similarity (41.7%) to Aur1 from Spathaspora passalidarum. A phenotypic assay with transformed S. cerevisiae and P. kudriavzevii indicated that two amino acid residues, Phe166 and Gly249, play crucial roles in the resistance to aureobasidin A, which is consistent with previous reports for other fungal Aur1s. The GenBank Accession No. for PkAUR1 is KP729614.

Structural Analysis for Building Structures reflecting Differential Column Shortening based upon Construction Sequence (시공 공정에 따른 건축 구조물의 구조해석 및 수직부재의 부등변형)

  • 조상규;이형우;최창식
    • Proceedings of the Computational Structural Engineering Institute Conference
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    • 1998.10a
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    • pp.333-341
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    • 1998
  • The general method of structural analysis for building structures has been based upon the assumption that all dead loads are imposed on a building simultaneously throughout the entire structure. In reality, buildings are built floor by floor or a few floors at a time. The construction dead load is applied gradually onto the structure as the structure is being erected. The prevailing commercial software for structural analysis used to date have resulted in the representation of inaccurate structural behaviors. The actual construction sequence and the loading of the structure ere not properly represented in the analysis. This paper identifies the source of the errors and develops the algorithm to account for the differential column shortening due to construction dead load based upon a given construction sequence

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Comparison and Sequence Analysis of the 3` - terminal Regions of RNA 1 of Barley Yellow Mosaic Virus

  • Lee, Kui-Jae
    • Plant Resources
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    • v.1 no.2
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    • pp.92-97
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    • 1998
  • An isolate of barley yellow mosaic virus(BaYMV-HN) obtained from Haenam, Korea was compared with two BaYMV strains. BaYMV-Ⅱ-1 from Japan and BaYMV-G from Germany. The sequence of the 3'-terminal 3817nucleotides[excluding the poly (A) tail] of RNA 1 of BaYMV-HN was determined to start within a long open reading frame coding for a part of the NIa-VPg polymerase(26 amino acids). NIa-Pro polymerase (343 amino acids), NIb polymerase(528 amino acids) and the entire capsid protein(297 amino acids), which is followed by a noncoding region(NCR) of 235 nucelotides. In the partial ORFs, BaYMV-HN shows higher sequence homology with BaYMV-Ⅱ-1(99.5%) than BaYMV-G(92.7%). The 3' non-coding regions of BaYMV-HN(235nt) shows higher nucleotide sequence homology with BaYMV-G(235nt)(99.6%) than BaYMV-Ⅱ-1(231nt)(97.0%). The 3' NIa-Pro protein sequence of BaYMV-HN shows higher amino acid sequence homology with BaYMV-Ⅱ-1(95.0%) than BaYMV-G(93.6%), but, NIb protein sequence of BaYMV-HN shows same all amino acid sequence. The capsid protein sequence of BaYMV-HN(297aa) shows same with BaYMV-Ⅱ-1, and shows higher nucleotide sequence homology with BaYMV-UK (from United Kingdom)(97.3%) than BaYMV-G(96.9%) and G2(96.9%). Difference of capsid protein amino acid were 0-9 between the Japan, United Kingdom and Germany and were 2-6 between all Korean isolates. Many of the amino acid differences are located in the N-terminal regions of the capsid proteins from 1 to 74 amino acid positions.

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Cloning and Sequence Analysis of Wild Argali ISG15 cDNA

  • Sun, Yanming;Chen, Kaili;Shen, Wen;Cui, Rupeng;Lu, Haifu
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.561-566
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    • 2014
  • The complete coding sequence of Wild Argali ISG15 cDNA was generated by rapid amplification of cDNA ends. The ISG15 cDNA was 642 bp with an open reading frame of 474 bp, which encoded a 17.47 kDa protein composed of 157 amino acids. Its amino acid sequence shared 97.9%, 80.8%, 91.4%, 94.3%, 78.3% identity with those of ISG15cDNA from Ovis aries (accession no. NM001009735.1), Capra hircus (accession no. HQ329186.1), Bos taurus (accession no. BC102318.1), Bubalus bubalis (accession no. HM543269.1), and Sus scrofa (accession no. EU647216.1), respectively. The entire coding sequence was inserted into the pET-28a vector and expressed in E. coli. The recombinant protein corresponded to the expected molecular mass of 25 kDa as judged by SDS-PAGE, and it was detected in the bacterial inclusion bodies. The expressed protein could be purified by $Ni^{2+}$ chelate affinity chromatography and the results from the lymphocyte proliferation test showed that the product could stimulate lymphocyte proliferation very well (p<0.05), which further confirmed its biological activity.

Identification of Positive and Negative Regulatory Elements of the Human Cytochrome P4501A2 (CYP1A2) Gene

  • Chung, Injae;Jeong, Choonsik;Jung, Kihwa;Bresnick, Edward
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.81-81
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    • 1997
  • We previously demonstrated an enhancer-like positive regulatory element within a 259-bp sequence (-2352 to-2094 bp) of the human CYP1A2 gene in HepG2 cells. Three protein binding sites were identified by DNase I footprint analyses within the 259-bp sequence: protected region A PRA ( -2283 to-2243 bp), PRB (-2218 to-2187 bp), and PRC (-2124 to-2098 bp) (I. Chung and E. Bresnick, Mol. Pharmacol. 47, 677-685, 1995). In the present study, the functional significance of those protected regions was examined. Transfection experiments with deletion and substitution mutants defined the PRB and PRC as containing positive and negative regulatory elements, respectively. Human breast carcinoma MCF-7 cells were cotransfected with a hepatocyte nuclear factor-1 (HNF-1) expression vector and CYP1A2 promoter-or thymidine kinase promoter-luciferase remoter gene constructs. HNF-1, which contributes to the liver specificity of genes, enhanced reporter gene activity in a PRC sequence-dependent manner. These results suggested that PRC could exist bound to a repressor which was displaceable by other transcription factors such as HNF-1. Results obtained by transfection of HepG2 hepatoma cells with various PRB substitution mutant-luciferase gene fusion constructs indicated that the entire sequence of PRB was necessary for promoter activity. Consequently, the regulation of CYP1A3 expression is very complex, requiring a number of both positive and negative regulatory factors.

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Cloning and Nucleotide Sequence of a cDNA Encoding the Rat Triosephosphate Isomerase

  • Lee, Kyunglim;Ryu, Jiwon
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.497-501
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    • 1996
  • A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1, 348-bp cDNA clone contains 24 bp $5^I$ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, $547bp 3^I$ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI CDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu13O, Tyr2O9, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.

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Complete genome sequence analysis Hosta virus X and comparison to other potexviruses

  • Park, M.H.;K.H. Ryu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.113.1-113
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    • 2003
  • A potexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants (Hosta spp.), and its entire genome RNA sequence was determined. in Korea using cDNA library and RACE methods. The genome of HVX encodes five open reading frames coding for viral replicase, triple gene block (TGB), and viral coat protein (CP) from the 5'to 3' ends, which is a typical genome structure of potexviruses. The 3-terminal region of the virus includes the TGBI (26 kDa), TGB2 (13 kDa), TGB3 (8 kDa), and 23 kDa coat protein (CP) and the 3-nontranslated region (NTR). The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100% and 98.9% identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50% nucleotide identical to other sequenced potexviruses. This is the first report of complete genome sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus.

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