• Title/Summary/Keyword: Recombinant vector

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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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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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Helper-Independent Live Recombinant Adenovirus Vector Expressing the Hemagglutinin-Esterase Membrane Glycoprotein

  • YOO, DONGWAN;ICK-DONG YOO;YOUNG-HO YOON;FRANK L GRAHAM;LORNE A. BABIUK
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.174-182
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    • 1992
  • The hemagglutinin-esterase glycoprotein (HE) gene of bovine coronavirus, coupled with a simian virus 40 early promoter and polyadenylation signal, was inserted into a human adenovirus transfer vector. The transfer vector was used to co-transfect 293 cells along with adenovirus genomic DNA. The hemagglutinin-esterase transcription unit was rescued into the adenovirus genome by homologous in vivo DNA recombination between the vector plasmid DNA and the adenovirus genomic DNA, and a recombinant adenovirus was isolated by several rounds of plaque assays. Thus the recombinant adenovirus carries the hemagglutinin-esterase gene in the early transcription region 3 (E3) of the adenovirus genome in the parallel orientation to the E3 transcription. The recombinant adenovirus synthesized the HE polypeptide in HeLa cells as demonstrated by immunoprecipitation with anti-coronavirus rabbit antisera. The recombinant HE polypeptide could be labelled by $[^3H]$glucosamine, demonstrating that the recombinant HE was glycosylated. Cells expressing the HE polypeptide exhibited hemadsorption activity when incubated with mouse erythrocytes. The HE was transported to the plasma membrane as shown by the cell surface immunofluorescence, indicating that the recombinant HE polypeptide retained its biological activities. Potential for the use of infectious recombinant adenovirus as a live virus-vectored vaccine candidate for bovine coronavirus disease is discussed.

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Influence of Immunity Induced at Priming Step on Mucosal Immunization of Heterologous Prime-Boost Regimens

  • Eo, Seong-Kug
    • IMMUNE NETWORK
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    • v.3 no.2
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    • pp.110-117
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    • 2003
  • Background: The usefulness of DNA vaccine at priming step of heterologous prime-boost vaccination led to DNA vaccine closer to practical reality. DNA vaccine priming followed by recombinant viral vector boosting via systemic route induces optimal systemic immunity but no mucosal immunity. Mucosal vaccination of the reversed protocol (recombinant viral vector priming-DNA vaccine boosting), however, can induce both maximal mucosal and systemic immunity. Here, we tried to address the reason why the mucosal protocol of prime-boost vaccination differs from that of systemic vaccination. Methods: To address the importance of primary immunity induced at priming step, mice were primed with different doses of DNA vaccine or coadministration of DNA vaccine plus mucosal adjuvant, and immunity including serum IgG and mucosal IgA was then determined following boosting with recombinant viral vector. Next, to assess influence of humoral pre-existing immunity on boosting $CD8^+$ T cell-mediated immunity, $CD8^+$ T cell-mediated immunity in B cell-deficient (${\mu}K/O$) mice immunized with prime-boost regimens was evaluated by CTL assay and $IFN-{\gamma}$-producing cells. Results: Immunity primed with recombinant viral vector was effectively boosted with DNA vaccine even 60 days later. In particular, animals primed by increasing doses of DNA vaccine or incorporating an adjuvant at priming step and boosted by recombinant viral vector elicited comparable responses to recombinant viral vector primed-DNA vaccine boosted group. Humoral pre-existing immunity was also unlikely to interfere the boosting effect of $CD8^+$ T cell-mediated immunity by recombinant viral vector. Conclusion: This report provides the important point that optimally primed responses should be considered in mucosal immunization of heterologous prime-boost regimens for inducing the effective boosting at both mucosal and systemic sites.

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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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 Bioluminescent Labelling Plasmid Vector for Bifidobacteria

  • Moon, Gi-Seong;Narbad, Arjan
    • Food Science of Animal Resources
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    • v.38 no.4
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    • pp.816-822
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    • 2018
  • Bifidobacterium is recognized as one of the most beneficial microorganisms in our gut. Many researches on bifidobacteria have been done to understand their roles in the gut. The objective of the present study was to develop a bioluminescent labelling plasmid vector for bifidobacteria to facilitate their visualization in vitro, in situ, and in vivo. A plasmid replicon (2.0 kb) of plasmid pFI2576 previously identified from B. longum FI10564 was amplified by PCR and cloned into pUC19 plasmid vector (2.68 kb). The cloned replicon was subcloned into pTG262 ($luc^+$) recombinant plasmid vector (7.4 kb) where a luciferase gene ($luc^+$) from pLuc2 (8.5 kb), an Escherichia coli and lactobacilli shuttle vector, was inserted into pTG262 plasmid vector. The final recombinant DNA, pTG262::pFI2576 rep ($luc^+$), was transferred into a B. catenulatum strain. This recombinant strain showed 3,024 relative luminescence units at $OD_{600}$ value of 0.352. Thus, this recombinant plasmid construct can be broadly used for labelling bifidobacteria.

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.

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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