• Title/Summary/Keyword: ie-1

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Analysis of Promoter Strength of Autographa californica Nuclear Polyhedrosis Virus IE1 Gene by Using Rreconmbinant Baculovirus

  • Cho, Eun-Sook;Park, Hae-Jin;Jin, Byung-Rae;Sohn, Hung-Dae;Kang, Seok-Woo;Yun, Eun-Young;Kim, Keun-Young;Je, Yeon-Ho;Kang, Seok-Kwon
    • Journal of Sericultural and Entomological Science
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    • v.41 no.2
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    • pp.102-107
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    • 1999
  • To analysis a promoter strength of Atographa californica nucler polyhedrosis virus (AcNPV) IE1 gene, an immediate viral gene, ${\beta}$-glactosidase gene as a reporter gene was introduced under the control of the IE1 promoter. The restriction fragment containing IE1 promoter and ${\beta}$-galctosidase gene from pAcIE1-gal were inserter into pBacPAK9 to yield transfer vector pAcNPV-IE1-gal. The pAcNPV-IE1-gal was cotransfected with AcNPV genomic DNA BacPAK6 into Sf9 cells to produce recombinant baculovirus AcNPV-IE1-gal. In addition, recombinant bacvulovirus AcNPV-gal, which express ${\beta}$-galac-tosidase under the control of the polyhedrin promoter, was constrer, was constructed to compared with AcNPV-IE1-gal. The recombinant viruses were respectively infected into Sf9 cells and characterized by the virus titer and expression of ${\beta}$-galactoxidase in Sf9 cells. The promoter strength of IE1 and polyhedrin promoters was determined by the amount of ${\beta}$-galactosidase secreted into medium by viral infection. The titer of AcNPV-IE1-Gal determined by plaque assays in Sf9 cells was similar to that of AcNPV-gal. However, expression level of ${\beta}$-galactosidase by AcNPV-IE1-gal was significantly lower than that by AcNPV-gal. In conclusion, promoter strength of IE1 was approximately 25-fold lower than that of polyhedrin.

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Mapping of Human Cytomegalovirus IE1 Responsive Elements in the c-jun Promoter (사람세포거대바이러스 (Human Cytomegalovirus)의 극초기항원-1 (Immediate Early-1, IE-1)에 반응하는 c-jun Promoter의 유전자 지도 분석)

  • Park, Chung-Gyu;Han, Tae-Hee;Kim, Dae-Joong;Kim, Jin-Hee;Hwang, Eung-Soo;Choi, Sung-Bae;Cha, Chang-Yong
    • The Journal of Korean Society of Virology
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    • v.28 no.3
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    • pp.267-274
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    • 1998
  • Human cytomegalovirus (HCMV) has the ability to activate the expression of many viral and cellular genes. Among various viral proteins, the immediate early proteins (IE1-72kDa, IE2-86kDa) have been known to be potent transactivators. The product of c-jun proto-oncogene is important in cell activation and differentiation. Here, we tried to find out if the IE could activate the c-jun promoter and also tried to identify the responsible sequence elements in the c-jun activation by IE1-72kDa. We found HCMV IE expression transactivated the c-jun promoter in human embryonal lung fibroblasts (HEL). The activation fold by IE1-72kDa, IE2-86kDa and IE2-55kDa was 23, 35, and 5, respectively. When the expression of each IE was combined, it showed synergism. Expression of (IE1-72kDa + IE2-86kDa) and (IE1-72kDa + IE2-86kDa + IE2-55kDa) resulted in 131 and 162 fold increase, respectively. The c-jun promoter region between -117 and -59 contains binding sites for the transcription factors Spl, CAAT, AP-l like (ATF/CREB), and MEF2. Transient expression assays were performed using various reporter plasmids containing the c-jun promoter-regulatory region linked to the luciferase gene and a plasmid expressing HCMV IE1 gene. Deletional and point mutational analysis showed that the sequence between -225 to -160 and the CTF binding site were involved in the up-regulation of c-jun promoter.

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Identification and Characterization of a Putative Baculoviral Transcriptional Factor IE-1 from Choristoneura fumiferana Granulovirus

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Merzouki, Abderrazzak;Guertin, Claude
    • BMB Reports
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    • v.35 no.6
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    • pp.553-561
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    • 2002
  • A gene that encodes a protein homologue to baculoviral IE-1 was identified and sequenced in the genome of the Choristoneura fumiferana granulovirus (ChfuGV). The gene has an 1278 nucleotide (nt) open-reading frame (ORF) that encodes 426 amino acids with an estimated molecular weight of 50.33 kDa. At the nucleotide level, several cis-acting regulatory elements were detected within the promoter region of the ie-1 gene of ChfuGV along with other studied granuloviruses (GVs). Two putative CCAAT elements were detected within the noncoding leader region of this gene; one was located on the opposite strand at -92 and the other at -420 nt from the putative start triplet. Two baculoviral late promoter motifs (TAAG) were also detected within the promoter region of the ie-1 gene of ChfuGV. A single polyadenylation signal, AATAAA, was located 18nt downstream of the putative translational stop codon of ie-1 from ChfuGV. At the protein level, the amino acid sequence data that was derived from the nucleotide sequence in ChfuGV IE-1 was compared to those of the Cydia pomonella granulovirus (CpGV), Xestia c-nigrum granulovirus (XcGV) and Plutella xylostella granulovirus (PxGV). The C-terminal regions of the granuloviral IE-1 sequences appeared to be more conserved when compared to the N-terminal regions. A domain, similar to the basic helix-loop-helix like (bHLH-like) domain in NPVs, was detected at the C-terminal region of IE-1 from ChfuGV (residues 387 to 414). A phylogenetic tree for baculoviral IE-1 was constructed using a maximum parsimony analysis. A phylogenetic estimation demonstrates that ChfuGV IE-1 is most closely related to that of CpGV.

Characterization of the molecular and biological properties between the equine herpesvirus type 1 immediate-early protein and the general transcription factor human TFIIB

  • Jang Hyung-Kwan
    • Korean Journal of Veterinary Service
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    • v.27 no.4
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    • pp.355-369
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    • 2004
  • The equine herpesvirus type 1 (EHV-1) immediate-early (IE) protein is a potent transactivator responsible for the activation of both early and late genes during the course of infection and is comprised of discrete functional domains that mediate its many functions. Interaction between trans activators such as the IE protein and various components of the RNA polymerase II transcription initiation machinery has been demonstrated to be critical for transactivation. In the present report, it is addressed the hypothesis that the IE protein interacts with various components of transcription machinery to mediate transactivation of target viral genes. In these studies, it is demonstrated that in vitro transcribed and translated IE protein interacts with TFIIB-agarose conjugate but not with TFIID-agarose conjugate. Additional immunoprecipitation studies using nuclear extracts derived from EHV-1 infected RK-13 cells confirmed that the IE protein interacts strongly with TFIIB, but fails to interact with TFIID. IR2, a truncated form of the IE protein lacking the potent transactivation domain and involved in the down-regulation of the IE gene, also interacted with TFIIB but not with TFIID. Studies were also performed to ascertain if particular TBP-associated factors (TAFs) could mediate IE or IR2 binding to TFIID. In vitro transcribed and translated TAF250 added to nuclear extracts generated from EHV-1 infected cells also failed to mediate an interaction between the IE protein or the IR2 protein and TFIID. This study demonstrated that the IE protein mediates transactivation of target viral genes by a mechanism that involves TFIIB. This is in contrast to mechanisms that have been proposed for both the herpes simplex virus ICP4 and VP16 protein which have been proposed to transactivate viral genes through interactions involving both TFIIB and TFIID. This study also intimates that IR2 mediate its repressive effects during the course of EHV-1 infection by a mechanism that involves sequestration of various transcription factors.

Mapping of the equine herpesvirus type 1 immediate-early protein interaction domain within the general transcription factor human TFIIB

  • Jang, Hyung-Kwan;Cho, Jeong-Gon;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.25 no.4
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    • pp.333-346
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    • 2002
  • We previously reported that the equine herpesvirus type 1(EHV-1) immediate-early protein(IE protein) physically interacts with the general transcription factor human TFIIB(Jang et al, J Virol 75:10219-10230, 2001). The interaction between the IE protein and TFIIB is necessary for the IE protein to efficiently transactivate the early TK and late IR5 EHV-1 promoters. A panel of deletion and truncation mutants of the TFIIB gene was constructed and employed in protein-binding assays to map the IE protein-binding domain within TFIIB. Evidence is presented that the first direct repeat of TFIIB interacts specifically with the EHV-1 IE protein.

Technical note: Estimation of Korean industry-average initiating event frequencies for use in probabilistic safety assessment

  • Kim, Dong-San;Park, Jin Hee;Lim, Ho-Gon
    • Nuclear Engineering and Technology
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    • v.52 no.1
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    • pp.211-221
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    • 2020
  • One fundamental element of probabilistic safety assessment (PSA) is the initiating event (IE) analysis. Since IE frequencies can change over time, time-trend analysis is required to obtain optimized IE frequencies. Accordingly, such time-trend analyses have been employed to estimate industry-average IE frequencies for use in the PSAs of U.S. nuclear power plants (NPPs); existing PSAs of Korean NPPs, however, neglect such analysis in the estimation of IE frequencies. This article therefore provides the method for and results of estimating Korean industry-average IE frequencies using time-trend analysis. It also examines the effects of the IE frequencies obtained from this study on risk insights by applying them to recently updated internal events Level 1 PSA models (at-power and shutdown) for an OPR-1000 plant. As a result, at-power core damage frequency decreased while shutdown core damage frequency increased, with the related contributions from each IE category changing accordingly. These results imply that the incorporation of time-trend analysis leads to different IE frequencies and resulting risk insights. The IE frequency distributions presented in this study can be used in future PSA updates for Korean NPPs, and should be further updated themselves by adding more recent data.

Construction of Stably Transformed Bm5 Cells by Using Autographa californica Nuclear Polyhedrosis Virus IE1 Gene

  • Cho, Eun-Sook;Jin, Byung-Rae;Sohn, Hung-Dae;Chol, Kwang-Ho;Kim, Soung-Ryul;Kang, Seok-Woo;Yun, Eun-Young;Kim, Sang-Hyun;Kim, Keun-Young
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.111-116
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    • 1998
  • To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

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IE Introduction History and Situation in Korea (우리나라 IE 도입과정과 현황)

  • Kim, Man-Sik
    • IE interfaces
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    • v.6 no.1
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    • pp.7-18
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    • 1993
  • Since scientific management techniques are introduced into Korea about 1958, the backwardness of Korea companies and the ignorant owners of the companies had been main obstacles in aboriginality of those techniques. With the activation of industry in 1970s, however, the adaptation of IE is also activated. This paper, although it may be slipshod, describes the present situation as well as the adaptation history of IE techniques. This is for the expected necessity of memorizing the introduction of IE in the future.

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Performance Enhancement of Combined-IMM/IE Filter for Tracking a Maneuvering Target (기동표적 추적을 위한 IMM/IE 혼합 필터의 성능개선)

  • Lim, Sang-Seok;Park, Jung-Ho
    • Journal of Advanced Navigation Technology
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    • v.5 no.1
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    • pp.74-84
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    • 2001
  • Recently a new algorithm which combines advantages of the IMM and IE methods has been suggested. The combined-IMM/IE algorithm could improve the performance to some extent. However, the problem of large increase of tracking error near the maneuver detection due to the sudden maneuver input has not been solved. In this paper, we propose two schemes which can resolve this limitations of combined-IMM/IE algorithm. For illustrations of the performance of the proposed methods. Monte-Carlo simulations are carried out and the results are analyzed.

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An IE-FFT Algorithm to Analyze PEC Objects for MFIE Formulation

  • Seo, Seung Mo
    • Journal of electromagnetic engineering and science
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    • v.19 no.1
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    • pp.6-12
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    • 2019
  • An IE-FFT algorithm is implemented and applied to the electromagnetic (EM) solution of perfect electric conducting (PEC) scattering problems. The solution of the method of moments (MoM), based on the magnetic field integral equation (MFIE), is obtained for PEC objects with closed surfaces. The IE-FFT algorithm uses a uniform Cartesian grid to apply a global fast Fourier transform (FFT), which leads to significantly reduce memory requirement and speed up CPU with an iterative solver. The IE-FFT algorithm utilizes two discretizations, one for the unknown induced surface current on the planar triangular patches of 3D arbitrary geometries and the other on a uniform Cartesian grid for interpolating the free-space Green's function. The uniform interpolation of the Green's functions allows for a global FFT for far-field interaction terms, and the near-field interaction terms should be adequately corrected. A 3D block-Toeplitz structure for the Lagrangian interpolation of the Green's function is proposed. The MFIE formulation with the IE-FFT algorithm, without the help of a preconditioner, is converged in certain iterations with a generalized minimal residual (GMRES) method. The complexity of the IE-FFT is found to be approximately $O(N^{1.5})$and $O(N^{1.5}logN)$ for memory requirements and CPU time, respectively.