• Title/Summary/Keyword: baculovirus expression vector system

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Efficacy of Gene Transfer of Recombinant Baculovirus Vector

  • Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2013.05a
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    • pp.1006-1008
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    • 2013
  • A novel recombinant baculovirus vector system containing coding genes for polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) was constructed. We applied this recombinant baculovirus vector into cells and murine tissues and compared efficacy of gene transfer and expression of this recombinant baculovirus vector system with control vector system. From this result, we confirmed that this novel recombinant baculovirus vector system was very effective than control vector system.

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Gene Transfer and Gene Expression of Novel Recombinant Baculovirus Vector System (새로운 재조합 베큘로바이러스벡터의 유전자전이와 유전자발현)

  • Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2013.10a
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    • pp.946-948
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    • 2013
  • Several baculovirus vector systems recombined with coding genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were applied into human foreskin fibroblast cells and compared the effects of gene transfer and gene expression of these recombinant baculovirus vector systems with control vector system. From this study, it showed that these novel recombinant baculovirus vector systems were superior efficacy to control vector system in view of gene transfer and gene expression.

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Efficacy of Gene Transfer and Expression of Novel Recombinant Baculovirus Vector (새로운 재조합 베큘로바이러스 벡터의 유전자 전달과 유전자 발현의 효과)

  • Kweon, Tae-Dong;Hong, Seong-Karp
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.8
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    • pp.2017-2022
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    • 2014
  • Novel baculovirus vector systems recombined with coding genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were applied into human foreskin fibroblast cells and compared the effects of gene transfer and gene expression of these recombinant baculovirus vector systems with control vector system. From this study, it showed that these novel recombinant baculovirus vector systems were superior efficacy to control vector system in view of gene transfer and gene expression.

Novel Construction of Recombinant Baculovirus Vector System (재조합 베큘로바이러스 벡터 시스템의 신 구축)

  • Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2012.10a
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    • pp.994-996
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    • 2012
  • We constructed novel recombinant baculovirus vector system. This vector system contained coding genes for polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). We compared efficacy and rate of expression of this novel recombinant baculovirus vector system with other control vector system. From this result, we confirmed that this novel recombinant baculovirus vector system was superior to other control vector system.

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Comparison of Recombinant Baculovirus Vector Systems and Control Vector System (재조합 베큘로바이러스벡터와 대조 벡터의 비교)

  • Kim, Ji-Young;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2015.05a
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    • pp.954-957
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    • 2015
  • A recombinant baculovirus vector systems were composed of genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). These recombinant baculovirus vector system were transfected into various cell lines and tissues and confirmed gene transfer and expression of these vector systems with only control vector system. From the result, gene transfer and gene expression of recombinant baculovirus vector systems were superior in terms of efficacy and safety than in the control vector system.

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Efficacy of Gene Transfer and Expression of Recombinanat Baculovirus Vector System (재조합 베큘로바이러스벡터의 유전자전달과 발현의 효과)

  • Sa, Young-Hee;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2014.05a
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    • pp.813-815
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    • 2014
  • Novel baculovirus vector systems including genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD) were constructed. These recombinant baculovirus vector systems were transfected into diverse cells of 293T, HepG2, HFF, and Hur7 cells and compared the effects of gene transfer and expression of these vector systems with control vector. From the result, we confirmed that these recombinant baculovirus vector systems were more excellent than control vector in efficacy of gene transfer and expression.

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Constructions of a Transfer Vector Containing the gX Signal Sequence of Pseudorabies Virus and a Recombinant Baculovirus

  • Lee, Hyung-Hoan;Kang, Hyun;Kim, Jung-Woo;Hong, Seung-Kuk;Kang, Bong-Joo;Song, Jae-Young
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.541-547
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    • 1999
  • Constructions of a transfer vector and a recombinant baculovirus using the thymidine kinase gene of the Herpes simplex virus type 1 strain F (HSV -1) were carried out. Newly cloned transfer vector, pHcgXIIIB, was constructed by insertion of the glycoprotein gX gene signal peptide sequence of Pseudorabies virus into the baculovirus vector pHcEV-IV. The gX sequence was inserted just downstream from the promoter for the polyhedrin gene of the Hyphantria cunea nuclear polyhedrosis virus (HcNPV). HSV-1 thymidine kinase(tk) gene (1.131 kb) was used as a candidate gene for transferring into the baculovirus expression system. The tk gene was inserted into a BamHI site downstream from the gX sequence-promoter for the polyhedrin gene in the pHcgXIIIB transfer vector and was transferred into the infectious lacZ-HcNPV expression vector. Recombinant virus was isolated and was named gX-TK-HcNPV. The recombinant virus produced a 45 kDa gX-TK fusion protein in Spodoptera frugiperda cells, which was confirmed by Western blot analysis. Microscopic examination of gX-TK-HcNPV-infected cells revealed normal multiplication. Fluorescent antibody staining indicated that the gX-TK fusion protein was present in the cytoplasm. These results indicated that the transfer vector successfully transferred the gX-tk gene into the baculovirus expression system.

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Expression of Human Stem Cell Factor with Recombinant Baculovirus in BmN Cell Line and Silkworm

  • Xijie, Guo;Yongfeng, Jin;Mingguan, Yang;Yaozhou, Zhang
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.51-56
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    • 2002
  • A recombinant transfer vector pBacSCF was constructed by inserting huamn stem cell factor (hSCF) cDNA into plasmid pBacPAK8. BmN cells were co-transfected with modified Bombyx mori, nuclear polyhedrosis virus (BmBacPAK) DNA and the recmbinant transfer vector to construct a recombinant baculovirus containing hSCE gene. DNA dot blotting and RNA dot blotting demonstrated that the hSCE gene was contained in the recombinant virus and transcribed. The recombinant baculovirus was infectious to BmN cells and to silkworm. SDS-PAGE analysis showed a specific band of expressed product in the extract of infected cells and in the heamolymph of infected larvae. Bioactivity of the recombinant hSCE was determined with W-1 cell line and MTT colorimetric method in synergy with interlukin-3 (IL-3). These results revealed that the hSCF gene was over-expressed in cultured cells and lavae of silkworm.

Construction of the Novel Baculovirus Transfer Vector Using the p10 Gene of BmNPV (BmNPV의 p10 유전자를 이용한 새로운 전이벡터 개발)

  • 강석우;진병래
    • Journal of Sericultural and Entomological Science
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    • v.39 no.2
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    • pp.180-185
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    • 1997
  • To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polygedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5' and 3' flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, $\beta$-galactosidase gene was inserted in the SmaI site of foreign gene cloning site of pBm10. The pBm10 containing $\beta$-galactosidase gene was cotranfected wth genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing $\beta$-galactosidase was also produced polygedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.

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Cloning of Major Capsid Protein Gene of Pseudorabies Virus and Expression by Baculovirus Vector System (Pseudorabies Virus의 Major Capsid Protein 유전자의 클론닝과 Baculovirus Vector System에 의한 발현)

  • An, Dong-Jun;Jun, Moo-Hyung;Song, Jae-Young;Park, Jong-Hyeon;Hyun, Bang-Hun;Chang, Kyung-Soo;An, Soo-Hwan
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.151-162
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    • 1996
  • Pseudorabies is caused by Pseudorabies virus (PRV: Aujeszky's disease virus) of Herpesviridae that is characterized by 100 to 150nm in size with a linear double-stranded DNA molecule with of approximately $90{\times}10^6Da$. This disease affects most of domestic animals such as swine, cattle, dog, sheep, cat, chicken, etc. causing high mortality and economic losses. In swine, young piglets show high mortality and pregnant sows, reproductive failures. However the adult swine reveals no clinical signs in general. But they become a carrier state and play an important role for propagation of the disease. In this study, the nucleotide sequence of major casid protein gene of PRV, Yangsan strain isolated from the diseased swine in Korea was analyzed, and the recombinant MCP was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. As result, in BamHI digestion, MCP gene locus of PRV YS strain showed different from that of Indiana S strain. The patterns of enzyme mapping were also found to be unidentical each other. The sequence of the MCP gene partially analyzed showed 98.09% identity to Indiana S strain. The expression of MCP in Sf-9 cell cotransfected by pVLMCP-44 baculovirus expression vector was characterized by Southern blot hybridization, immunofluoresent and immunocytochemical tests, SDS-PAGE and Western blotting. The rMCP with M.W. 142kDa was most effectively expressed in Sf-9 cells at the 3-4th days post inoculation of the recombinant baculovirus by 2 moi.

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