• Title/Summary/Keyword: expression vectors

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Analysis of Efficiency of Recombinant pOPINEneo-3C-GFP Vector with p53 Tumor Suppression Gene Inserted (p53 암억제 유전자가 삽입된 재조합 pOPINEneo-3C-GFP 벡터의 효율 분석)

  • 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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    • 2019.05a
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    • pp.533-536
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    • 2019
  • Recombinant baculoviruses are widely used to express heterologous genes in cultured insect cells. Recombinant baculoviruses can serve as gene-transfer vectors for expression of recombinant proteins in a wide range of mammalian cell types. Baculovirus system has significant benefits in view of safety, large-scale, and high level of gene expression. In this study, baculoviral vectors which were reconstructed from pOPINEneo-3C-GFP vector, were recombined with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), and p53 with NcoI and XhoI. 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. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

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Embodied Approach to the Concept of Vector and its Application

  • Cho, Han Hyuk;Noh, Chang Kyun;Choi, In Yong
    • Research in Mathematical Education
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    • v.18 no.4
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    • pp.289-305
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    • 2014
  • The current mathematical education calls for a learning environment from the constructionism perspective that actively creates mathematical objects. This research first analyzes JavaMAL's expression 'move' that enables students to express the agent's behavior constructively before they learn vector as a formal concept. Since expression 'move' is based on a coordinate, it naturally corresponds with the expression of vectors used in school mathematics and lets students take an embodied approach to the concept of vector. Furthermore, as a design tool, expression 'move' can be used in various activities that include vector structure. This research studies the educational significance entailed in JavaMAL's expression 'move'.

Immune gene expression and protection effect against VHSV by injection of interferon regulatory factor 10 in zebrafish (Danio rerio) (제브라피쉬 interferon regulatory factor 10의 주사에 따른 면역 유전자 발현과 VHSV에 대한 방어 효과)

  • Kim, Hye Ji;Kim, Jin Young;Park, Jong Bin;Lee, Ji Hyun;Park, Jeong Su;Kim, Hyoung Jun;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.23-29
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    • 2021
  • Interferon regulatory factors (IRFs) are a family of transcription factors essential to the control of antiviral immune response, cell growth, differentiation and apoptosis. IRF10 of zebrafish (Danio rerio) was negative regulation of the interferonΦ1 and 3 response in vitro. In this study, we analyze the induction of in vivo immune response activation from the IRF10 gene of zebrafish and the protective effect against VHSV. As the results, the group inoculated with IRF10 expression vectors, there was no expression of IFNΦ1, suggestion that IRF10 may function as a negative regulator of IRF3, which binds to the IFNΦ1 promoter. And other types of interferon genes (IFNΦ2-4) are thought to have been activated, inducing to the expression of pro-inflammatory cytokine and Mx genes. As the results of challenge test performed at 14 days after inoculation of the expression vectors, the maximum survival rate [50% (1㎍ DNA) and 42.5% (10㎍ DNA)] for IRF10 group were recorded. Meanwhile, the survival rates of pcDNA3.1 and PBS as the control groups were 10% and 15%, respectively. This study suggests that the possibility that activation of IRF10 molecule could be exploited as a VHS control method.

Construction of a Temperature Controlled Expression Ve e tor in Saccharumy ces cerevisiae (Saccharomyces cerevisiae를 이용한 온도조절형 발현 Vector의 개발)

  • 최진옥;황용일
    • Microbiology and Biotechnology Letters
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    • v.21 no.3
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    • pp.214-220
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    • 1993
  • The mating type a of yeast, Saccharomyces cerevisiae mutant with hmla2-102 and sir3-8ts was changed to type alpha by changing the growth temperature from 25C to 35C. A temperature-sensitive expression vector system was constructed using mating factor alpha1 (Mfalpha1) gene encoding alpha factor which is expressed in the type alpha cells. Vectors with different copy numbers were constructed by joining the promoter and pre or prepro-secretion single sequence of Mfalpha1 to promoterless PHO5' gene as a reporter gene.

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A plasmid vector faciliting gene expression in both yeast and mammalian cells

  • Lee, Tae-Ho
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.149-151
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    • 1997
  • A plasmid vector with combined features of yeast shuttle vector and mammalian expression vector was constructed to facilitate expression of cloned gene in both cell-types. All necessary elements required for plasmid maintenance and selection in E. coli, yeast and mammalian cells were size-economically arranged in this plasmid. The numan cytomegalovirus (CMV) immediate early promoter and yeast GAL1 promoter were sequentially placed in front of the gene to be expressed. The synthetic splicing donor and acceptor sequences were inserted into the immediate upstream and downstream of the GAL1 promotor, allowing the CMV promotor to direct the expression of a given gene in mammalian cell environment by splicing out the interfering GAL1 promotor sequence. When the resulting vector containing LacZ as a gene was introduced into yeast and mammalian cells, both cells efficiently produced .betha.-galactosidase, dimonstrating its dual host usage.

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Soluble Expression of Human Angiostatin and Endostatin by Maltose Binding Protein (MBP) Fusion in E. coli (Maltose Binding Protein 융합단백질에 의한 인간유래의 앤지오스타틴과 앤도스타틴의 대장균에서 수용성 단백질발현)

  • Paek, Seon-Yeol;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.B
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    • pp.59-63
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    • 2008
  • Rapid production of therapeutic proteins such as angiostatin and endostatin angiogenic inhibititors has been highly demanded for cancer treatment. In this regard, recombinant human angiostatin and endostatin were successfully expressed as soluble forms by maltose binding protein (MBP)-mediated fusion expression in Escherichia coli. PCR amplified, angiostatin and endostatin genes from human placenta cDNA library were inserted into an expression vector pMAL-c2e to construct prokaryotic expression vectors, pMAL-c2e/AS and pMAL-c2e/ES, respectively. Recombinant angiostatin and endostatin were efficiently expressed in E. coli origami (DE3) after IPTG induction and protein expression were confirmed by SDS-PAGE analyses. The expressed recombinant proteins were purified near homogenity using an amylose affinty column chromatography. In contrast that previous E. coli expressions were all insoluble, our results first time demonstrated that MBP fused human angiostatin and endostatin were soluble in E. coli.

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Cloning and Expression of Human Immunodeficiency Virus-1 Epitopes in Escherichia coli (대장균에서 사람의 면역결핍 바이러스-1 epitopes 클로닝과 발현에 대한 연구)

  • 유향숙;장원희;박희동;현상원;남상욱;이영익
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.1-7
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    • 1991
  • Human immunodeficiency virus type 1 (HIV-1) causes a deadly infectious disease, Acquired Immunodeficiency Syndrome (ADIS). As a first step to develop a reliable and fast diagnostic procedure for HIV-1 infection, we cloned various immunodominant epitopes of HIV-1 in bacterial expression vectors containing tac or trp promoter. While the protein level of direct expression of gp160 was low, trp E fused gp120, gp41 and p17-p24 were produced at high levels (15-30% of total bacterial proteins) in E. coli. Since gp120 and gp41 contain relatively conserved regions which can react with antibodies in the plasma from most of HIV-1 infected individuals, these expression clones were used for large preparations of HIV-1 antigens.

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