• 제목/요약/키워드: Recombinant vector

검색결과 717건 처리시간 0.034초

Efficacy of Gene Transfer of Recombinant Baculovirus Vector

  • Sa, Young-Hee;Hong, Seong-Karp
    • 한국정보통신학회:학술대회논문집
    • /
    • 한국정보통신학회 2013년도 춘계학술대회
    • /
    • pp.1006-1008
    • /
    • 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.

  • PDF

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

  • 사영희;홍성갑
    • 한국정보통신학회:학술대회논문집
    • /
    • 한국정보통신학회 2012년도 추계학술대회
    • /
    • pp.994-996
    • /
    • 2012
  • 본 연구실에서는 새로운 베큘로바이러스 벡터 시스템을 구축하였다. 즉 본 벡터 시스템은 polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD)을 코딩하는 유전자들로 구성 되어있다. 이렇게 새로이 제작된 베큘로바이러스 벡터 시스템과 대조군의 벡터 시스템과 효율과 발현율을 비교하였다. 그 결과 본 연구실에서 제작된 베큘로바이러스 벡터 시스템이 다른 대조군의 벡터 시스템에 비해 효과적임을 확인할 수 있었다.

  • PDF

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
    • /
    • 제2권3호
    • /
    • pp.174-182
    • /
    • 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.

  • PDF

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

  • Eo, Seong-Kug
    • IMMUNE NETWORK
    • /
    • 제3권2호
    • /
    • pp.110-117
    • /
    • 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.

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

  • 김지영;홍성갑
    • 한국정보통신학회:학술대회논문집
    • /
    • 한국정보통신학회 2015년도 춘계학술대회
    • /
    • pp.954-957
    • /
    • 2015
  • polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 유전자로 구성된 재조합 베큘로바이러스를 제작하였다. 본 재조합 베큘로바이러스 시스템은 여러가지 세포주와 여러 가지 조직에 감염하여 시험하였고 재조합된 유전자의 전달과 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구의 결과로 제작된 재조합 베큘로바이러스 시스템은 유전자의 전달과 발현에 있어서 대조 벡터시스템 보다 효능과 안전성면에서 우수하였다.

  • PDF

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
    • /
    • 제4권1호
    • /
    • pp.51-56
    • /
    • 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
    • 한국축산식품학회지
    • /
    • 제38권4호
    • /
    • pp.816-822
    • /
    • 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.

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

  • 사영희;홍성갑
    • 한국정보통신학회:학술대회논문집
    • /
    • 한국정보통신학회 2013년도 추계학술대회
    • /
    • pp.946-948
    • /
    • 2013
  • 베큘로바이러스 시스템이 제조되었는데 이것은 polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 유전자를 재조합한 것이다. 본 재조합벡터 시스템은 인간 섬유아세포에 적용하여 시험하였고 재조합된 유전자의 전이와 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구로부터 새롭게 제작된 본 베큘로바이러스 시스템이 유전자의 전이와 유전자 발현 면에서 대조 벡터시스템 보다 고효율을 나타내었다.

  • PDF

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

  • 권태동;홍성갑
    • 한국정보통신학회논문지
    • /
    • 제18권8호
    • /
    • pp.2017-2022
    • /
    • 2014
  • 폴리히드론 프로모터, 수포성구내염 바이러스G, 폴리A, 사이토메가바이러스 프로모터, 강화녹색형광단백질, 단백질전달부위 유전자 등을 포함한 새로운 베큘로바이러스 시스템이 제조되었다. 본 재조합벡터 시스템은 인간 섬유아세포에 적용하여 시험하였고 재조합된 유전자의 전달과 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구로부터 새롭게 제작된 본 베큘로바이러스 시스템이 유전자의 전이와 유전자 발현 면에서 대조 벡터시스템 보다 고효율을 나타내었다.

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

  • 김지영;홍성갑
    • 한국정보통신학회:학술대회논문집
    • /
    • 한국정보통신학회 2014년도 추계학술대회
    • /
    • pp.977-980
    • /
    • 2014
  • polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 유전자가 포함된 재조합 베큘로바이러스를 구축하였다. 본 재조합 베큘로바이러스 시스템은 인간 섬유아세포와 여러 가지 조직에 감염하여 시험하였고 재조합된 유전자의 전달과 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구의 결과로 제작된 재조합 베큘로바이러스 시스템은 유전자의 전달과 발현에 있어서 대조 벡터시스템 보다 더욱 효과적이고 안전적이었다.

  • PDF