• Title/Summary/Keyword: Characteristic sequence

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Cloning, Expression, and Characterization of DNA Polymerase from Hyperthermophilic Bacterium Aquifex pyrophilus

  • Choi, Jeong-Jin;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1022-1030
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    • 2004
  • The gene encoding Aquifex pyrophilus (Apy) DNA polymerase was cloned and sequenced. The Apy DNA polymerase gene consists of 1,725 bp coding for a protein with 574 amino acid residues. The deduced amino acid sequence of Apy DNA. polymerase showed a high sequence homology to Escherichia coli DNA polymerase I-like DNA polymerases. It was deduced by amino acid sequence alignment that Apy DNA polymerase, like the Klenow fragment, has only the two domains, the $3'{\rightarrow}5'$ exonuclease domain and the $5'{\rightarrow}3'$ polymerase domain, containing the characteristic motifs. The Apy DNA polymerase gene was expressed under the control of T7lac promoter on the expression vector pET-22b(+) in E. coli. The expressed enzyme was purified by heat treatment, and Cibacron blue 3GA and $UNO^{TM}$ Q column chromatographies. The optimum pH of the purified enzyme was 7.5, and the optimal concentrations of KCl and $Mg^{2+}$ were 20 mM and 3 mM, respectively. Apy DNA polymerase contained a double strand-dependent $3'{\rightarrow}5'$ proofreading exonuclease activity, but lacked any detectable $5'{\rightarrow}3'$ exonuclease activity, which is consistent with its amino acid sequence. The somewhat lower thermostability of Apy DNA polymerase than the growth temperature of A. pyrophilus was analyzed by the comparison of amino acid composition and pressure effect.

Determining 3D-shape of specular objects by using an encoded grid pattern light source

  • Ye, Xiongying;Fujimura, Sadao
    • 제어로봇시스템학회:학술대회논문집
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    • 1991.10b
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    • pp.1758-1763
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    • 1991
  • This paper describes a new method to determine the 3D-shape of objects consisting of specular planar surfaces. This method exploits a light source which is made of a diffuse plane with a grid pattern encoded in an M-sequence and uses a single image of the light source reflected by the objects to acquiring orientations and positions of the surfaces of the objects. When grid lines of the light source are reflected by a specular planar surface and perspectively projected on an image plane, a set of lines vanishing at a point are obtained on the image plane. The orientation of the specular planar surface is determined by using the vanishing point, and the position is determined by using the correspondence between lines on the image and lines on the light source, which is obtained by employing a characteristic regularity of the M-sequence. Before the vanishing points are calculated, the lines on the image are classified and correlated with the surfaces of objects by using slopes and positions of the lines and the regularity of the M-sequence. This method requires only a single image.

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The 52 kD Protein Gene of Odontoglossum Ringspot Virus Containing RNA-Dependent RNA Polymerase Motifs and Comparisons with Other Tobamoviruses

  • Park, Won-Mok
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.129-136
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    • 1995
  • Complementary DNA of the genomic RNA of odontoglossum ringspot virus Cymbidium strain (ORSV-Cy) was synthesized from polyadenylated viral RNA and cloned. Selected clones containing the viral RNA-dependent RNA polymerase gene of the virus has been sequenced by automated sequencing system. The complete nucleotide sequence of an open reading frame is 1377 base pairs in length, and encodes a protein of 458 amino acids about 52, 334 D. The 52 kD protein of ORSV shares four sequence motifs characteristic of viral RNA-dependent RNA polymerase. Comparison of the ORSV 52 kD protein sequence with that of other five viruses in tobamovirus group showed 76.0 to 60.7% homologies at the amino acid level and the conservation of the four motifs betwen the viruses.

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A study on the Drilling according to the Stacking Sequence of the CFRP (탄소섬유강화플라스틱의 적층구성에 따른 드릴 가공에 관한연구)

  • Kim, S.B.;Shin, H.G.;Kim, T.Y.
    • Journal of the Korean Society of Mechanical Technology
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    • v.13 no.2
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    • pp.1-7
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    • 2011
  • The CFRP composite has a lot of merits such as mechanical characteristic, light and thermal resistance. For these merits, CFRP is applied to so many industrial area. In order to use the composite materials in the aircraft structures or machine elements, accurate surfaces for bearing mounting or joints must be provided, which require precise, machining. In this study, the specimens differentiating the stacking sequence of 5kinds were used. When drilling the carbon fiber reinforced plastics, it was checked on whether the stacking sequence reached any effect on the cutting force. Also relationship between the drill diameter is examined from the drilling experiment, which is the drilling of Fabric, Unidirectional specimen with ∅6mm, ∅10mm, ∅12mm cemented carbide drill. Considering cutting force and drilling diameter, the results are analyzed.

Optimal stacking sequence of laminated anisotropic cylindrical panel using genetic algorithm

  • Alibeigloo, A.;Shakeri, M.;Morowa, A.
    • Structural Engineering and Mechanics
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    • v.25 no.6
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    • pp.637-652
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    • 2007
  • This paper presents stacking sequence optimization of laminated angle-ply cylindrical panel based on natural frequency. Finite element method (FEM) is used to obtain the vibration characteristic of an anisotropic panel using the first order shear deformation theory(FSDT) and genetic algorithm (GA) is used to obtain the optimal stacking sequence of the layers. Cylindrical panel has finite length and arbitrary boundary conditions. The thicknesses of the layers are assumed constant and their angles are specified as design variables. The effect of the number of plies and boundary conditions in the fitness function is considered. Numerical examples are presented for four, six and eight layered anisotropic cylindrical panels.

An Approach for a Substitution Matrix Based on Protein Blocks and Physicochemical Properties of Amino Acids through PCA

  • You, Youngki;Jang, Inhwan;Lee, Kyungro;Kim, Heonjoo;Lee, Kwanhee
    • Interdisciplinary Bio Central
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    • v.6 no.4
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    • pp.3.1-3.10
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    • 2014
  • Amino acid substitution matrices are essential tools for protein sequence analysis, homology sequence search in protein databases and multiple sequence alignment. The PAM matrix was the first widely used amino acid substitution matrix. The BLOSUM series then succeeded the PAM matrix. Most substitution matrixes were developed by using the statistical frequency of substitution between each amino acid at blocks representing groups of protein families or related proteins. However, substitution of amino acids is based on the similarity of physiochemical properties of each amino acid. In this study, a new approach was used to obtain major physiochemical properties in multiple sequence alignment. Frequency of amino acid substitution in multiple sequence alignment database and selected attributes of amino acids in physiochemical properties database were merged. This merged data showed the major physiochemical properties through principle components analysis. Using factor analysis, these four principle components were interpreted as flexibility of electronic movement, polarity, negative charge and structural flexibility. Applying these four components, BAPS was constructed and validated for accuracy. When comparing receiver operated characteristic ($ROC_{50}$) values, BAPS scored slightly lower than BLOSUM and PAM. However, when evaluating for accuracy by comparing results from multiple sequence alignment with the structural alignment results of two test data sets with known three-dimensional structure in the homologous structure alignment database, the result of the test for BAPS was comparatively equivalent or better than results for prior matrices including PAM, Gonnet, Identity and Genetic code matrix.

MSC-based Test-case Generation Module for Railway Signaling Software Testing (철도신호 소프트웨어 테스팅을 위한 MSC 기반 테스트케이스 생성 모듈)

  • Hwang, Jong-Gyu;Baek, Jong-Hyun
    • The Transactions of the Korean Institute of Electrical Engineers P
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    • v.64 no.3
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    • pp.138-142
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    • 2015
  • Most of the existing studies on functional safety testing for the railway signaling system software have focused on verifying the functional safety through the monitoring of internal memory embedded railway signaling system. However, the railway signaling system is one of the typical embedded control system in the railway sector, and the embedded software has a characteristic of generating an appropriate outputs through the combination of internal processing in consideration of the current internal status and external input. Therefore, the test approach of using the interface communication channel can be effective way for the functional testing for railway signaling system software in consideration of these characteristic. Since a communication interface specification of the railway signal system has a the properties of the sequence input and output signals, test-case for software testing is the most effective methodology by MSC (Message Sequence Chart) language, one of the graphic language. The MSC-based test-case generating methodology for signaling system software was proposed in this paper.

Phylogenetic Analysis of Hepatitis B Virus Genome Isolated from Korean Patient Serum

  • Kim, Seon-Young;Kang, Hyen-Sam;Kim, Yeon-Soo
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.823-828
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    • 2000
  • The complete nucleotide sequence of hepatitis B virus DNA isolated from Korean patient serum was determined and characterized, and its phylogenetic relation was then investigated. The viral genome was 3,215 base pairs long and included four well known open reading frames (i.e. surface antigens, core antigens, X protein and DNA polymerase). The sequence of the surface antigen showed that the HBV genome under investigation, designated HBV 315, was characteristic of subtype adr. A phylogenetic analysis using the total genome sequence revealed that HBV315 was grouped into genomic group C together with isolates from Japan, China, Thailand, Polynesia, and New Caledonia. The mean percent similarity between HBV315 and other HBV isolates in genomic group C was 97.25%, and that with other genomic groups ranged from 86.16% to 91.25%. The predicted amino acid sequences of HBV315 were compared with two closely related subtype adr isolates, M38636 and D12980. The results showed that the X gene product was identical in the three strains, while there were significant amino acid sequence differences between HBV315 and M38636 in the Pre-S1 and Pre-S2 regions.

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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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Isolation and characterization of a noval membrane-bound cytochrome $C_{553}$ from the strictly anaerobic phototroph, heliobacillus mobilis

  • Lee, Woo-Yiel;Bla;Kim, Seung-Ho
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.206-212
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    • 1997
  • Heliobacillus mobilis is a strictly anaerobic Gram-positive bacterium which contains a primitive Photosystem I-type reaction center. The membrane-bound cytochrome $C_{553}$ from the heliobacterium suggested to be the immediate electron donor to the photooxidized pigment (P798+) has been isolated and characterized. The heme protein was visualized as a major component with an apparent molecular size of 17kDa in TMBZ-staining analysis of the membrane preparation and showed characteristic $\alpha$ (552.5 nm), $\beta$ (522nm), and Soret absorption (416 nm) peaks of a typical reduced c-type cytochrome in the partially purified sample. The internal 43 amino acid sequence of the electron donor was obtained by chemical agent and protease treatments followed by N-terminal sequencing of the resulting fragments. The internal sequence carries lots of lysine residues and a Cys-X-X-Cys-His sequence motif which are the characteristics of typical c-type cytochromes. The analysis of the sequence by FAST or FASTA program, however, did not show any significant similarity to other known heme proteins.

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