• Title/Summary/Keyword: baculovirus expression

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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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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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Generation of Baculovirus Expression Vector Using Detective Autographa California Nuclear Polyhedrosis Virus Genome Maintained in Escherichia coli for $Occ^{+}$ Virus Production

  • Je, Yeon-Ho;Chang, Jin-Hee;Roh, Jong-Yul;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.155-160
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    • 2001
  • We have generated a novel baculovirus genome which can be maintained in Escherichia coli that facilitates the rapid and efficient generation of recombinant baculovirus expression vectors. To make $Occ^{+}$ recombinant expression vectors, polyhedrin gene under the control of p10 promoter was inserted to bAcGOZA and this genome was designated bApGOZA. As in bAcGOZA, bApGOZA lacks a portion of the essential ORF1629 gene, but includes a mini-F replicon and selectable kanamycin-resistance marker, This occasion-producing activity of bApGOZA can be used very conveniently for its oral infectivity to insect larvae in mass production of foreign protein and insecticides.

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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.

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.

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 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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Expression of Immunologically Active Porcine Recombinant TGF-${\beta}1$ Precursor Protein in Baculovirus System

  • Lim, Hyun;Kim, Pyeung-Hyeun;Chun, Gie-Taek;Choi, Eui-Yul;Yie, Se-Won
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.341-346
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    • 1997
  • In order to express recombinant porcine TGF-${\beta}1$ protein in a baculovirus expression system the entire TGF-${\beta}1$gene containing extra amino acids at the N terminus was cloned into pFBa and pFBb of the Bac-To-$Bac^{TM}$ baculovirus expression system. One of the clones contained 106 extra amino acids and was designated pFBa-106 TGF-${\beta}1$, and the other had 28 extra amino acids and was designated pFBb-28 TGF-${\beta}1$. The orientation of the gene was identified with restriction enzyme mapping and PCR with internal TGF-${\beta}1$ primers. Sf-9 cells were infected at a m.o.i. of 10 by the recombinant viruses generated from the two expected sizes of 55 kD and 46.4kD. these precursor forms of TGF-${\beta}1$ with a polyclonal antibody against human TGF-${\beta}1$. No mature form of TGF-${\beta}1$ protein was detected on SDS gels and an immunoblot indicated that TGF-${\beta}1$ precursor is not properlu processed in insect cells.

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High-efficient Expression of Porcine IL-2 with Recombinant Baculovirus Infected Silkworm, Bombyx mori

  • Inumaru, Shigeki;KokuHo, Takehiro;Yada, Takashi;Kiuchi, Makoto;Miyazawa, Mitsusuhiro
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.146-149
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    • 2000
  • Biologically active porcine Interleukin-2(poIL-2) was produced from in vitro and in viva baculovirus expression system, namely the Autographa californica nuclear polyhedrosis virus (ACNPV)-cell culture system and the Hybrid nucler polthedrosis virus (HyNPV)-sillkworm larva system. The concentration of the recombinant poIL-2(rpoIL-2) in the larvae hemolymph was 1 to 3 mg/mL, which was about 7 to 20 times those of the cell culture systems. The level of this expression efficiency is equal to that with transgenic livestock, secretion products in milk.

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