• Title/Summary/Keyword: gp64

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Oligomerization of the substitution mutants of autographa californica nuclear polyhedrosis Virus (AcNPV) gp64 glycoprotein

  • Kim, Ki-Nam;Poo, Ha-Ryoung;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.35 no.1
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    • pp.72-77
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    • 1997
  • The baculovirus gp64 glycoprotein is a major component of the envelope protein of budded virus (BV). It has been shown that the gp64 glycoprotein plays an essential role in the infection process, especialy fusion between virus envelope and cellular endosomic membrane. Recently we reported optimal conditions required for gp64-mediated membrane fusion in pGP64 DNA transfected Spodoptera frugiperda (Sf9) cells (H. J. Kim and J. M. Yang, Jour, Microbiology, 34.7-14). In order to investigate the role of hydrophobicity within the fusion domain of the gp64 glycoprotein for membrane fusion, 13 mutants which have substitution mutation within hydrophobic region I were constructed by PCR-derived site-derected mutagenesis. Each mutated gp64 glycoproteins was transiently expressed by transfecting plasmid DNA into Spodoptera frugiperda (Sf9) cells. Oligomerization of the transisently expressed gp64 glycoproteins was a nalysed by running them on SDS-polyacrylamide gel electrophoresis under non-reducing condition followed by immunoblotting. All of the mutant gp64 glycoproteins expect cysteine-228 were able to form trimers. These results suggest that hydrophobic region I of the gp64 may not be responsible for the oligomerization of the gp64 glycoprotein.

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Characterization of gp64 Gene of Bombyx mori Nucleopolyhedrovirus and Development of a Transient Expression Vector (누에 핵다각체병 바이러스 헤 gp64 유전자의 특성조사 및 transient 발현 벡터 개발)

  • 김미향;최재영;우수동;이해광;제연호
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.18-24
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    • 2001
  • Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

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Analysis of fusogenic activity of autographa californica nuclear polyhedrosis virus (Ac NPV) gp64 envelope glycoprotein

  • Kim, Hee-Jin;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.7-14
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    • 1996
  • Teh baculovirus gp64 glycoprotein is a major component of the envelope of budded virus (BV) and has been shown that it plays an essential role in the infection process, especially virus-cell membrane fusion. We have cloned Autographa californica Nuclear Polyhedrosis Virus (AcNPV) gp64 protein were examined for membrane fusion activity by using a synchtium formation assay under various conditions. The optimal conditions required for inducing membrane fusion are 1) form pH 4.0 to 4.8 2) 15 min exposure of cells to acidic pH 3) at least 1 .mu.g of gp64 cloned plasmid DNA per 3 * 10$^{6}$ cells 4) and an exposure of cells to acidic pH at 72 h post-transfection. In order to investigate the role of hydrophobicity of the gp64 glycoprotein for the membrane fusion, the two leucine residues (amino acid position at 229 and 230) within hydrophobic region I were substituted to alanine by PCR-derived site-directed mutagenisis and the membrane fusion activity of the mutant was anlaysed. The gp64 glycoprotein carrying double alamine substitution mutation showed no significant difference in fusion activity. This result suggested that minor changes in hydrophobicity at the amino acid position 229 and 230 does not affect the acid-induced membrane fusion activity of the gp64 glycoprotein.

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Expression of Human Thrombopoietin in Insect Cells by Polyhedrin-gp64 Dual Promoter-Based Baculovirus Vector System

  • Koh, Yeo-Wook;Park, Sang-Kyu;Kim, Kun-Soo;Kim, Seong-Ryong;Lee, Jeong-Kug;Yang, Jai-Myung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.318-322
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    • 1999
  • A new baculovirus transfer vector (pPGP404) was constructed to increase the expression level of human thrombopoietin (hTPO) in insect cells. In pPGP404, hTPO was cloned next to the AcNPV polyhedrin-gp64 dual promoter and the leader sequence of hTPO was substituted with that of gp64. A recombinant baculovirus, AcPGP404, was constructed by using pPGP404 as a transfer vector. hTPO was expressed in AcPGP404-infected TN5 cells and it was observed that the expression levels of hTPO in TN5 cells increased three-fold ($6.0 {\mu}g/ml^{-1}$) compared to the level expressed under the control of the polyhedrin single promoter. These results indicate that the polyhedrin-gp64 dual promoter system would be useful for expression in large quantities of recombinant proteins in insect cells.

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Petrological characteristics on stone resources of granites in the Pocheon-Euijeongbu area (포천-의정부지역 화강암류 석재자원의 암석학적 특성연구)

  • 윤현수
    • The Journal of the Petrological Society of Korea
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    • v.6 no.1
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    • pp.34-44
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    • 1997
  • The Jurassic granites, commercially called Yangu stone in the Pocheon-Euijeongbu area, have generally compact and coarse-grained textures, which could be classified into two types; grey granite(Gg) and light pink granite(Gp). Specific gravity, absorption ratio and prosity of Gg and Gp in physical property are 2.64 and 2.61, 0.32 % and 0.44 %, 0.86% and 1.13 %, respectively. These higher values of two latters of Gp than those of Gg are due to the more abundant microcracks in Gp. Compressive strength og Gg than those of Gg are due to more abundant microcracks in Gp. Compressive strength og Gg and Gp are 1,726 kg/cm2 and 1,717 kg/cm, respectively and bestrength has a positive proportion with Qz+Af+Pl(quartz+alkali feldspar+plagioclase) modes without trending with Bt+Ac(biotite+accessories). Tensile strength has the positive proportions with Qz+Af+Pl and Bt+Ac. While Bt+Ac has a negative trend with abrasive hardness, Qz+A+Pl shows a positive one. These may suggest Qz+Af+Pl mainly affects on strenghts potentional dimension stone than Gp.

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SOME RESULTS ON AN INTUITIONISTIC FUZZY TOPOLOGICAL SPACE

  • Min, Kyung-Ho;Min, Won Keun;Park, Chun-Kee
    • Korean Journal of Mathematics
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    • v.14 no.1
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    • pp.57-64
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    • 2006
  • In this paper, we introduce the concepts of $r$-closure and $r$-interior defined by intuitionistic gradation of openness. We also introduce the concepts of $r$-gp-maps, weakly $r$-gp-maps, and obtain some characterizations in terms of $r$-closure and $r$-interior operators.

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Improvement of Genetic Programming Based Nonlinear Regression Using ADF and Application for Prediction MOS of Wind Speed (ADF를 사용한 유전프로그래밍 기반 비선형 회귀분석 기법 개선 및 풍속 예보 보정 응용)

  • Oh, Seungchul;Seo, Kisung
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.64 no.12
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    • pp.1748-1755
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    • 2015
  • A linear regression is widely used for prediction problem, but it is hard to manage an irregular nature of nonlinear system. Although nonlinear regression methods have been adopted, most of them are only fit to low and limited structure problem with small number of independent variables. However, real-world problem, such as weather prediction required complex nonlinear regression with large number of variables. GP(Genetic Programming) based evolutionary nonlinear regression method is an efficient approach to attach the challenging problem. This paper introduces the improvement of an GP based nonlinear regression method using ADF(Automatically Defined Function). It is believed ADFs allow the evolution of modular solutions and, consequently, improve the performance of the GP technique. The suggested ADF based GP nonlinear regression methods are compared with UM, MLR, and previous GP method for 3 days prediction of wind speed using MOS(Model Output Statistics) for partial South Korean regions. The UM and KLAPS data of 2007-2009, 2011-2013 years are used for experimentation.

Expression and Characterization of Human Immunodeficiency Virus-1 Oligomerized gp140 Protein in Mammalian Cells (포유동물 세포에서 Human Immunodeficiency Virus-1의 Oligomeric gp140 단백의 발현 및 특성)

  • Kim, Eun-Ok;Kim, Eun;Kim, Hyun-Soo;Shin, Kwang-Soon;Kim, Chul-Joong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.55-64
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    • 2002
  • The envelope glycoprotein of HIV-1 forms an oligomeric complex resulting in playing a role to induce neutralizing antibody and cell-mediate immune responses. The oligomer exists as a trimer of gp120-gp41 heterodimer which mediates HIV-1 attachment and fusion. We made a cDNA clone of gp140 consisting of gp120 and ectodomain of gp41 from the primary African isolate. To express the oligomeric gp140 in mammalian cells, we adopted the Semliki Forest virus (SFV) based expression system. The oligomeric gp140 in the secretory form was expressed and purified from the cell culture supernatant and characterized. The antibody inducing activity of the purified gp140 was also examined in mice inoculation.

Evolutionary Generation Based Color Detection Technique for Object Identification in Degraded Robot Vision (저하된 로봇 비전에서의 물체 인식을 위한 진화적 생성 기반의 컬러 검출 기법)

  • Kim, Kyoungtae;Seo, Kisung
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.64 no.7
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    • pp.1040-1046
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    • 2015
  • This paper introduces GP(Genetic Programming) based color detection model for an object detection of humanoid robot vision. Existing color detection methods have used linear/nonlinear transformation of RGB color-model. However, most of cases have difficulties to classify colors satisfactory because of interference of among color channels and susceptibility for illumination variation. Especially, they are outstanding in degraded images from robot vision. To solve these problems, we propose illumination robust and non-parametric multi-colors detection model using evolution of GP. The proposed method is compared to the existing color-models for various environments in robot vision for real humanoid Nao.

Overexpression of a delayed early gene hlg1 of temperate mycobacteriophage L1 is lethal to both M. smegmatis and E. coli

  • Chattoraj, Partho;Ganguly, Tridib;Nandy, Ranjan Kumar;Sau, Subrata
    • BMB Reports
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    • v.41 no.5
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    • pp.363-368
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    • 2008
  • Two genes of temperate mycobacteriophage L5, namely, gp63 and gp64, were hypothesized to be toxic to M. smegmatis. An identical L5 gp64 ortholog (designated hlg1) was cloned from homoimmune mycobacteriophage L1 and characterized at length here. As expected, hlg1 affected the growth of M. smegmatis when overexpressed from a resident plasmid. HLG1 (the protein encoded by hlg1) in fact caused growth retardation of M. smegmatis and the region encompassing its 57-114 C-terminal amino acid residues was found indispensable for its growthretardation activity. Both nucleic acid and protein biosynthesis were severely impaired in M. smegmatis expressing HLG1. Interestingly, HLG1 also affected E. coli almost similarly. This putative delayed early lipoprotein did not participate in the lytic growth of L1.