• Title/Summary/Keyword: Recombinant baculovirus

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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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Efficacy of Recombinant Baculovirus Vector Reconstructed in pcDNA3.1 Vector (pcDNA3.1 벡터에서 재구성된 재조합 Baculovirus 벡터의 효능)

  • Sa, Young-Hee;Choi, Chang-Shik;Lee, Ki Hwan;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2018.10a
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    • pp.444-447
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    • 2018
  • Baculovirus expression systems have many known advantages including fast and cost-effective methods to generate large amounts of recombinant proteins in comparison to bacterial expression systems, particularly those requiring complex post-translational modifications. Especially, recombinant baculoviruses can transfer their vectors and express their recombinant proteins in a wide range of mammalian cell types. In this study, baculoviral vectors which were reconstructed from pcDNA3.1 vector, were recombined with cytomegalovirus (CMV) promoter,uroplakin II promoter, polyhedron promoter, vesicular stomatitis virus G (VSVG), enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector.

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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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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 a Novel Recombinant Baculovirus Producing Polyhedra with a Bacillus thuringiensis Cry1Ac Crystal Protein

  • Je, Yeon-Ho;Jin, Byung-Rae;Roh, Jong-Yul;Chang, Jin-Hee;Kang, Seok-Kwon
    • The Journal of Korean Society of Virology
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    • v.29 no.3
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    • pp.145-153
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    • 1999
  • We have now constructed a novel recombinant baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV) producing polyhedra with Bacillus thuringiensis (Bt) CryIAc crystal protein. The recombinant polyhedra produced by the recombinant baculovirus, Btrus, in insect cells was characterized. The recombinant baculovirus has two independent transcription units in opposite orientations with two promoters, p10 or polyhedrin gene promoter each initiating transcription of either native polyhedrin or fusion protein with polyhedrin and Bt Cry1Ac crystal protein. Surprisingly, this recombinant baculovirus stably produced recombinant polyhedra which were nearly similar to those of wild-type AcNPV. The immunogold staining experiment showed that the recombinant polyhedra were assembled with polyhedrin and Bt Cry1Ac crystal protein, and contained virus particles. Insecticidal toxicity of recombinant polyhedra of Btrus to the fall webworm, Hyphantria cunea, was strikingly improved in comparison with the wild-type AcNPV.

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Expression of Recombinant Human Bone morphogenetic protein 2 (hBMP2) in Insect cells

  • Kim, Seong-Wan;Kim, Seong-Ryul;Park, Seung Won;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.34 no.1
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    • pp.1-5
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    • 2017
  • Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

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.

Phosphorylation of the Nucleocapsid Protein of Bovine Coronavirus Expressed with a Recombinant Baculovirus Vector

  • Yoo, dongwan;Graham-J.Cox
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.122-128
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    • 1992
  • Post-translational modifications of the nucleocapsid protein of bovine coronavirus (Quebec strain) were investigated. Coronavirions were radiolabelled in vivo with inorganic $[^{32}P]$orthophosphate and analysed by SDS-PAGE, followed by autoradiography. A single polypeptide with a migration rate of 55 KDa was identified by metabolic phosphate labelling, demonstrating that the nucleocapsid protein of bovine coronavirus was a phosphoprotein. A gene encoding the nucleocapsid protein was inserted immediately downstream from the polyhedrin promoter of Autographa californica nuclear polyhedrosis baculovirus. Spodoptera frugiperda cells infected with this recombinant baculovirus synthesized a 55 KDa polypeptide, as demonstrated by immunoprecipitation with anti-nucleocapsid monoclonal antibody. The recombinant nucleocapsid protein synthesized in Spodoptera cells could also be labelled by $[^{32}P]$orthophosphate. Phosphoamino acid analysis showed that both serine and threonine residues were phosphorylated in authentic, as well as in recombinant nucleocapsid proteins, with a relative phosphorylation ratio of 7:3. Our studies demonstrated that the nucleocapsid protein of bovine coronavirus was a serine and threonine-phosphorylated protein and that Spodoptera insect cells were able to properly phosphorylate the relevant foreign proteins.

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Effective Expression of Recombinant Baculovirus Vector Systems (재조합 베큘로바이러스벡터의 효과적 발현)

  • Kim, Ji-Young;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2014.10a
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    • pp.977-980
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    • 2014
  • A 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 human foreskin fibroblast cells and various tissues and investigated gene transfer and expression of these vector systems with control vectors. From the study, these recombinant baculovirus vector systems were more effective and safe than control vector in view of gene transfer and expression.

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