• 제목/요약/키워드: Homology

검색결과 1,595건 처리시간 0.045초

cDNA Cloning and Expression of Human Rotavirus Outer Capsid Protein VP7 in Insect Cells

  • KANG, DU KYUNG;KI WAN KIM;PYEUNG-HYUN KIM;SEUNG YONG SEOUNG;YONG HEE KIM;ICK CHAN KWON;SEO YOUNG JEONG;EUI-YEOL CHOI;KYUNG MEE LEE
    • Journal of Microbiology and Biotechnology
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    • 제8권4호
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    • pp.369-377
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    • 1998
  • Rotavirus is a major cause of severe gastroenteritis in young children and animals throughout the world. The VP7 of rotavirus is thought to induce the synthesis of neutralizing antibodies and to be responsible for determining viral serotypes. The cDNA coding for the VP7 capsid protein of human rotavirus, obtained from Korean patients (HRV-Y14), was cloned and its nucleotide sequence was determined. Comparative analysis of the nucleotide sequences between VP7 of Y14 and that of other foreign isolates showed $92.7~95.2\%$ homology to G1 serotypes (RV-4, KU, K8, WA), $74.2\%$ homolgy to G2 serotype HU-5, $76.4\%$ homology to G3 serotype SA-11, and $77.6\%$ homology to G4 serotype A01321. These data suggest that HRV-Y14 can be classified as a G1 serotype. cDNA coding for VP7 of HRV-YI4 was subcloned into the baculovirus vector and the VP7 glycoprotein was expressed in insect cells. The expressed proteins in Sf9 cell extract and tissue culture fluid were separated on SDS-PAGE, and Western blot analysis with monoclonal antibody raised against the synthetic peptide containing 21 amino acids within the VP7 conserved region was performed. The molecular weight of recombinant VP7 was estimated to be 36 kDa which is about the same size as the native VP7. Addition of tunicamycin in the culture media caused a reduction of the molecular weight of the recombinant VP7 indicating that the expressed protein was glycosylated.

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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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    • 제1권2호
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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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COLOCALIZATION OF LOCAL HOMOLOGY MODULES

  • Rezaei, Shahram
    • 대한수학회보
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    • 제57권1호
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    • pp.167-177
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    • 2020
  • Let I be an ideal of Noetherian local ring (R, m) and M an artinian R-module. In this paper, we study colocalization of local homology modules. In fact we give Colocal-global Principle for the artinianness and minimaxness of local homology modules, which is a dual case of Local-global Principle for the finiteness of local cohomology modules. We define the representation dimension rI (M) of M and the artinianness dimension aI (M) of M relative to I by rI (M) = inf{i ∈ ℕ0 : HIi (M) is not representable}, and aI (M) = inf{i ∈ ℕ0 : HIi (M) is not artinian} and we will prove that i) aI (M) = rI (M) = inf{rIR𝖕 (𝖕M) : 𝖕 ∈ Spec(R)} ≥ inf{aIR𝖕 (𝖕M) : 𝖕 ∈ Spec(R)}, ii) inf{i ∈ ℕ0 : HIi (M) is not minimax} = inf{rIR𝖕 (𝖕M) : 𝖕 ∈ Spec(R) ∖ {𝔪}}. Also, we define the upper representation dimension RI (M) of M relative to I by RI (M) = sup{i ∈ ℕ0 : HIi (M) is not representable}, and we will show that i) sup{i ∈ ℕ0 : HIi (M) ≠ 0} = sup{i ∈ ℕ0 : HIi (M) is not artinian} = sup{RIR𝖕 (𝖕M) : 𝖕 ∈ Spec(R)}, ii) sup{i ∈ ℕ0 : HIi (M) is not finitely generated} = sup{i ∈ ℕ0 : HIi (M) is not minimax} = sup{RIR𝖕 (𝖕M) : 𝖕 ∈ Spec(R) ∖ {𝔪}}.

한국에서 분리된 IHNV-PRT의 G protein의 유전자 클로닝과 염기서열 분석 (Molecular Cloning and Nucleotide Sequence of the G protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus)

  • 김영조;허강준;박정우;박정문
    • 미생물학회지
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    • 제35권3호
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    • pp.226-230
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    • 1999
  • 무지개송어 등의 연어과 어류에서 발생하는 전염성 조혈기괴사증 바이러스의 국내분리주인 IHNV-PRT 의 특성을 규명하기 위하여 IHNV-PRT 의 당단백질인 G를 암호화하고 있는 유전자의 일부를 PCR로 증폭한 후 cDNA를 클로닝하여 이들의 염기서열을 분석하였다. 이 PCR product 는 442 bp의 크기이었다. IHNV-PRT 의 G의 염기서열을 IHNV 의 다른 strain 과 비교 분석한 결과 IHNV-RB-76, IHNV-RB, IHNV-LR-73, IHNV-K, IHNV-WRAC, IHNV-SRCV, IHNV-CoI-85들의 G와 각각 95,94,94,94,93,93%의 상동성을 보였다. 그러나 넙치로부터 분리된 fish rhabdovirus 인 hirame rhabdovirus 의 G와는 81%의 사동성을 보였다. 이 결과로부터 IHNV 의 G 유전자는 비록 HRV 의 G 유전자와 유사성은 높지 않지만, IHNV 의 strain 에 관계없이 유사하다는 것을 알 수 있다.

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Comparative Analysis of Nucleotide Sequence and Codon Usage of Arylphorin Gene Cloned from Four Silk-Producing Insects and Their Molicular Phylogenetics

  • Lee, Sang-Mong;Hwang, Jae-Sam;Lee, Jin-Sung;Goo, Tae-Won;Kwon, O-Yu;Kim, Ho-Rak
    • Journal of Life Science
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    • 제9권1호
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    • pp.84-89
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    • 1999
  • To determine phylogenetic relatedness of four silk-producing silkmoths (B. mori, B. mandarina, A. yamamai and A. pernyi), internal coding region of arylphorin which is a storage protein in hemolymph protein of insects were amplified by polymerase chain reaction and then sequenced and compared each other. The nucleotide composition was biased toward adenine and thymine(59% A+T) and a strong bias for use of C in the third position of codons was found for Phe and Tyr. Together TTC(Phe) and TAC(Tyr) account for about 16.8% (10 for TTC and 8 for TAC) of all codon usage. The nucleotide similarity of arylphorin gene from B. mori showed 99%, 98% and 97% homology with those of B. mandarina, A. yamamai and A. pernyi, respectively. Also, the nucleotide sequence of arylphorin gene from B. mandarina showed 98% and 97% homology with those of A. yamamai and A.pernyi, respectively. Between A. yamamai and A. pernyi, the sequence homology was 97%. The deduced amino acid sequences in B. mori, B. mandarina and A. yamamai showed almost 99% homology. Although the aryphorin gene provided insufficient variability among the four insect species, A UPGMA tree is generated that supported the monophyly of silk-producing insects, with M. sexta placed basal to it. It is suggest that silk-producing insects have a close relationship and a homogeneous genetic background from comparison with those of other insects.

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Toward the Virtual Screening of α-Glucosidase Inhibitors with the Homology-Modeled Protein Structure

  • Park, Jung-Hum;Ko, Sung-Min;Park, Hwang-Seo
    • Bulletin of the Korean Chemical Society
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    • 제29권5호
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    • pp.921-927
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    • 2008
  • Discovery of $\alpha$-glucosidase inhibitors has been actively pursued with the aim to develop therapeutics for the treatment of diabetes and the other carbohydrate mediated diseases. As a method for the discovery of new novel inhibitors of $\alpha$-glucosidase, we have addressed the performance of the computer-aided drug design protocol involving the homology modeling of $\alpha$-glucosidase and the structure-based virtual screening with the two docking tools: FlexX and the automated and improved AutoDock implementing the effects of ligand solvation in the scoring function. The homology modeling of $\alpha$-glucosidase from baker’s yeast provides a high-quality 3-D structure enabling the structure-based inhibitor design. Of the two docking programs under consideration, AutoDock is found to be more accurate than FlexX in terms of scoring putative ligands to the extent of 5-fold enhancement of hit rate in database screening when 1% of database coverage is used as a cutoff. A detailed binding mode analysis of the known inhibitors shows that they can be stabilized in the active site of $\alpha$- glucosidase through the simultaneous establishment of the multiple hydrogen bond and hydrophobic interactions. The present study demonstrates the usefulness of the automated AutoDock program with the improved scoring function as a docking tool for virtual screening of new $\alpha$-glucosidase inhibitors as well as for binding mode analysis to elucidate the activities of known inhibitors.

호몰로지 조건을 이용하여 충격과 마모를 고려한 원자로 핵연료봉 지지격자의 최적설계 (Optimization of a Nuclear Fuel Spacer Grid Using Considering Impact and Wear with Homology Constraints)

  • 이현아;김종기;송기남;박경진
    • 한국전산구조공학회:학술대회논문집
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    • 한국전산구조공학회 2007년도 정기 학술대회 논문집
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    • pp.145-150
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    • 2007
  • The spacer grid set is a component in the nuclear fuel assembly. The set supports the fuel rods saftely. Therefore, the spacer gl1d set should have sufficient strength for the external impact forces. The fretting wear occurs between the spring of the fuel rod and the spacer grid due to tile flow-induced vibration. The conceptual design of the spacer grid set is performed based on the Independence Axiom of axiomatic design. Two functional requirements are defined and corresponding design parameters are selected. The overall flow of the design is defined according to the application of axiomatic design. The design for the impact load is carried out by using nonlinear dynamic analysis to determine the length of the dimple. Topology optimization is carried out to determine a new configuration of the spring. The fretting wear is reduced by shape optimization using the homology theory. In the design to reduce the fretting wear, the deformed shape of the spring should be the same as that of the fuel rod. This condition is transformed to a function and considered as a constraint in the shape optimization process. The fretting wear is expected to be reduced due to the homology constraint. The objective function is minimizing the maximum stress to allow a slight plastic deformation. Shape optimization results are confirmed through nonlinear static analysis because the contact area becomes wider.

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Homology Modeling and Molecular Docking Study of Translationally Controlled Tumor Protein and Artemisinin

  • Chae, Jin-Sun;Choi, In-Hee;Kim, Choon-Mi
    • Archives of Pharmacal Research
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    • 제29권1호
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    • pp.50-58
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    • 2006
  • Translationally controlled tumor protein (TCTP), also known as histamine releasing factor (HRF), is found abundantly in different eukaryotic cell types. The sequence homology of TCTP between different species is very high, belonging to the MSS4/DSS4 superfamily of proteins. TCTP is involved in both cell growth and human late allergy reaction, as well as having a calcium binding property; however, its primary biological functions remain to be clearly elucidated. In regard to many possible functions, the TCTP of Plasmodium falciparum (Pf) is known to bind with an antimalarial agent, artemisinin, which is activated by heme. It is assumed that the endoperoxide-bridge of artemisinin is opened up by heme to form a free radical, which then eventually alkylates, probably to the Cys14 of PfTCTP. Study of the docking of artemisinin with heme, and subsequently with PfTCTP, was carried out to verify the above hypothesis on the basis of structural interactions. The three dimensional (3D) structure of PfTCTP was built by homology modeling, using the NMR structure of the TCTP of Schizosaccharomyces pombe as a template. The quality of the model was examined based on its secondary structure and biological function, as well as with the use of structure evaluating programs. The interactions between artemisinin, heme and PfTCTP were then studied using the docking program, FlexiDock. The center of the peroxide bond of artemisinin and the Fe of heme were docked within a short distance of $2.6{\AA}$, implying the strong possibility of an interaction between the two molecules, as proposed. When the activated form of artemisinin was docked on the PfTCTP, the C4-radical of the drug faced towards the sulfur of Cys14 within a distance of $2.48{\AA}$, again suggesting the possibility of alkylation having occurred. These results confirm the proposed mechanism of the antimalarial effect of artemisinin, which will provide a reliable method for establishing the mechanism of its biological activity using a molecular modeling study.

Comparative Homology Modeling and Ligand Docking Study of Human Catechol-O-Methyltransferase for Antiparkinson Drug Design

  • Lee, Jee-Young;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • 제26권11호
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    • pp.1695-1700
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    • 2005
  • Catechol-O-methyltransferase (COMT, EC 2.1.1.6) is an S-adenosylmethionine (SAM, AdoMet) dependent methyltransferase, and is related to the functions of the neurotransmitters in various mental processes, such as Parkinson’s disease. COMT inhibitors represent a new class of antiparkinson drugs, when they are coadministered with levodopa. Based on x-ray structure of rat COMT (rCOMT), the three dimensional structure of human COMT (hCOMT) was constructed by comparative homology modeling using MODELLER. The catalytic site of these two proteins showed subtle differences, but these differences are important to determine the characterization of COMT inhibitor. Ligand docking study is carried out for complex of hCOMT and COMT inhibitors using AutoDock. Among fifteen inhibitors chosen from world patent, nine models were energetically favorable. The average value of heavy atomic RMSD was 1.5 $\AA$. Analysis of ligand-protein binding model implies that Arg201 on hCOMT plays important roles in the interactions with COMT inhibitors. This study may give insight to develop new ways of antiparkinson drug.

The road less traveled: strategies to enhance the frequency of homology-directed repair (HDR) for increased efficiency of CRISPR/Cas-mediated transgenesis

  • Devkota, Sushil
    • BMB Reports
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    • 제51권9호
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    • pp.437-443
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    • 2018
  • Non-homologous end joining (NHEJ), and to a lesser extent, the error-free pathway known as homology-directed repair (HDR) are cellular mechanisms for recovery from double-strand DNA breaks (DSB) induced by RNA-guided programmable nuclease CRISPR/Cas. Since NHEJ is equivalent to using a duck tape to stick two pieces of metals together, the outcome of this repair mechanism is prone to error. Any out-of-frame mutations or premature stop codons resulting from NHEJ repair mechanism are extremely handy for loss-of-function studies. Substitution of a mutation on the genome with the correct exogenous repair DNA requires coordination via an error-free HDR, for targeted transgenesis. However, several practical limitations exist in harnessing the potential of HDR to replace a faulty mutation for therapeutic purposes in all cell types and more so in somatic cells. In germ cells after the DSB, copying occurs from the homologous chromosome, which increases the chances of incorporation of exogenous DNA with some degree of homology into the genome compared with somatic cells where copying from the identical sister chromatid is always preferred. This review summarizes several strategies that have been implemented to increase the frequency of HDR with a focus on somatic cells. It also highlights the limitations of this technology in gene therapy and suggests specific solutions to circumvent those barriers.