• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.70-71
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• 2005

본 연구는 1세포기 닭 수정란에 retrovirus vector (RSV-GFP)를 도입하여 외래유전자의 핵 전이 효율을 높이고자 하였다. 실험은 polybrene과 retrovirus 혼합물을 1세포기 또는 배반엽 단계의 수정란 세포질에 미세주입하고 배양 3 또는 4일차에 GFP의 발현 양상들을관찰하였다. 실험의 결과는 배반엽 수정란에서 GFP발현을 관찰할 수 있었으나, 1세포기 수정란에서는 GFP의 발현을 관찰할 수 없었다. 연구결과는 형질전환닭 생산에 있어서 가장 효율적인 방법은 배반엽 단계에 retrovirus를 미세주입하는 방법임을 보여주고 있다.

• Journal of fish pathology
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• v.16 no.2
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• pp.69-73
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• 2003

In this study, GFP reporter gene was transfected into a fibroblast cell line RTG-2 using a polycationic transfection reagent (Superfect) and showed a successful expression of GFP. The transfection efficiency by Superfect was compared to the commonly used transfection method, i.e. DNA-calcium phosphate coprecipitatlon. Transfection by Superfect was more effective than calcium phosphate coprecipltation method (frequency of cell expressing orr was 11.3% and 3.5%, respectively). The optimal expression of GFP and {\beta}-galactosidase was observed when $5-6\;{\mu}{\ell}$ of Superfect per ${\mu}g$ DNA was used for transfcction, 1:5-6 ratio between DNA(${\mu}g$) and Superfect ($\mu\ell$).

• Animal cells and systems
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• v.10 no.2
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• pp.73-77
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• 2006

Caspase-11 has been known as a dual regulator of apoptosis and inflammatory response. An unusual feature of caspase-11 is that its expression is induced by apoptotic or proinflammatory stimuli. Utilizing these unusual features of caspase-11, we have developed a simple and sensitive assay method to screen pro- or anti-apoptotic/inflammatory molecules. To develop this assay method, we generated a reporter construct where GFP expression is regulated by caspase-11 promoter. When several types of cultured cells were transfected with this reporter construct and subsequently treated with various apoptotic or proinflammatory molecules, expression of GFP by the activation of caspase-11 promoter was easily detected by fluorescence microscopy or spectrofluorometry. In addition, a reduction of the GFP fluorescence was detected when an agent reported to suppress caspase-11 induction was applied. These results suggest that our reporter system can be used to screen pro- or anti-apoptotic/inflammatory molecules.

• Journal of fish pathology
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• v.33 no.2
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• pp.163-169
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• 2020

Snakehead rhabdovirus (SHRV) is different from other fish novirhabdoviruses such as viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), and hirame rhabdovirus (HIRRV) in that it replicates at high temperatures. Therefore, the delivery of foreign proteins to fish living at high water temperature would be possible by using recombinant SHRVs. In the present study, to evaluate the possible use of SHRV as a vehicle for foreign proteins delivery, we generated a recombinant SHRV that contains an enhanced-GFP (eGFP) gene between nucleoprotein (N) and phosphoprotein (P) genes (rSHRV-A-eGFP), and another recombinant SHRV expressing two heterologous genes by inserting an eGFP gene between N and P genes, and mCherry gene between P and M genes (rSHRV-AeGFP-BmCherry). Epithelioma papulosum cyprini (EPC) cells infected with the recombinant SHRVs showed strong fluorescence(s), suggesting the possible availability of recombinant SHRVs for the development of combined vaccines by expressing multiple foreign antigens.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.149-152
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• 2000

In recent years, various artificial molecular photodiode have been fabricated by mimicking the electron transport function of biological photosynthesis. And now, we have been investigated the protein-organic hetero thin film photodiode using GFP as an sensitizer based on the redox potential difference of functional molecules. In this paper, shows molecular photodiode consisting of green fluorescence protein(GFP). viologen and TCNQ. The TCNQ and viologen were deposited onto ITO coated glass by LB technique. And GFP molecule was adsorption onto the viologen LB film surface by self-assembly method. Finally, The Al deposition onto GFP/viologen/TCNQ film surface was performed to make a top electrode. As a result, The MIM(metal/Insulator/Metal) structured device was constructed. The input light of 460nm wavelength was generated by the xenon lamp system, and then the photocurrent produced from the molecular device was detected through a current-voltage(I-V) measuring unit (SMU Model 236, Keithley, USA). An artificial molecular photodiode using protein(GFP)-adsorbed hetero-LB film is presented as a model system for the bioelectronic device based on the biomimesis.

• Journal of fish pathology
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• v.24 no.3
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• pp.297-305
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• 2011

A trap identification system for isolating functional sequences to allow the constitutive expression of foreign protein from Edwardsiella tarda was developed. Using the green fluorescent protein (GFP) reporter-based trap system, various functional sequences to drive heterologous expression of the GFP were selectable in Escherichia coli host. However from the bioinformatic sequence analysis, all the segments predicted as regulatory regions were not native promoters actually existing upstream of endogenous E. tarda genes. Instead, a number of non-authentic sequences, possibly resulted from the random shuffling and/or intermolecular ligation were also proven to be able to display a potent GFP expression in the recombinant E. coli. Further analysis with selected clones showed that both authentic and non-authentic sequences could function in as a constitutive promoter, leading quite a consistent and stable GFP expression after repetitive subcultures. Microscopic examination also confirmed the uniform pattern of GFP expression in every host bacterium. Semi-quantitative assay of GFP showed that there was no clear relationship between expression levels and organizational features of the promoters trapped. Functional promoter-like elements achieved in the present study could be a good starting material for multivalent genetic engineering of E. tarda in order to produce recombinant vaccines in a cost-effective fashion.

• Korean Journal of Microbiology
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• v.47 no.2
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• pp.174-178
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• 2011

In Escherichia coli, DcuS/R two-component system controls fumarate import and utilization related gene expression. To understand the dynamic response of the bacterium DcuS/R two-component system with respect to fumarate concentrations, DcuS/R induced dctA promoter was integrated with GFP reporter protein. Expression monitoring study using recombinant strain showed that dctA promoter was upregulated with 1 mM of fumarate in M9 minimal medium.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.10
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• pp.1367-1372
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• 2000

The present study was conducted to evaluate the efficiency of transgenic rabbit production by DNA microinjection using EGFP (Enhanced Green Fluorescent Protein) gene. In this experiment EGFP coding sequences fused to CMV promoter were microinjected into rabbit one-cell embryos, and then GFP expression and gene integration were evaluated in preimplantation embryos and fetuses recovered on day 15 of pregnancy to determine efficiency of transgenic rabbit production. Effect of DNA concentration was also tested on development in vitro following microinjection and transgene integration in fetuses. Development of embryos in vitro was decreased by DNA microinjection, but the rates of pregnancy and implantation were not significantly affected by microinjection. As development progressed in vitro percentage of GFP expression in rabbit embryos was decreased, resulting GFP expression detected in 37.5% of blastocysts. The efficiencies for production of transgenic fetuses were 4.0% and 7.6%, respectively, when $10ng/{\mu}l$ and $20ng/{\mu}l$ of DNA concentration were microinjected. Transgenic fetuses were confirmed by GFP expression and PCR analysis of fetus genomic DNA. These results indicated that DNA microinjection itself damaged embryo development and DNA concentration affected the efficiency of transgenic rabbit production.

• Journal of Microbiology and Biotechnology
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• v.12 no.1
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• pp.65-70
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• 2002

A TGF-${\beta}1$/GFP monomeric fusion protein was cloned from pPK9A and pGFP-Cl plasmid by PCR amplification. The fusion protein was expressed in a $Bac-To-Bac^{TM}$ baculovirus expression system. A 45 kDa fusion protein was purified using an Ni-NTA column with 300 mM imidazol from a cell lysate infected with recombinant viruses for 72 h post-infection. The fusion protein cross-reacted with the commercial $TGF-{\beta}1$ polyclonal Ab as well as Ab raised against a precursor, monomeric $TGF-{\beta}1$, and GFP. The binding activity of the fusion protein with a $TGF-{\beta}1$ receptor was examined. Fluorescence was observed in Mv1Lu cells, yet not in insect cells treated with the fusion protein. No fluorescence was detected in Mv1Lu cells incubated with the fusion protein treated with Ab prior to the binding reaction, or with GFP alone, thereby indicating that the binding of the fusion protein was specific to $TGF-{\beta}1$ with a receptor.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.677-680
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• 2007

To analyze a cotG-based Bacillus subtilis spore display system directly, $GFP_{UV}$ was expressed on the surface of Bacillus subtilis spores. When $GFP_{UV}$ was fused to the C-terminal of the cotG structural gene and expressed, the existence of a $CotG-GFP_{UV}$ fusion protein on the B. subtilis spore was confirmed by flow cytometry confocal microscopic analysis. When the cotG anchoring motif was deleted, no fluorescence emission was observed under flow cytometry and confocal microscopic analysis from the purified spore, confirming the essential role of CotG as an anchoring motif. This $GFP_{UV}$ displaying spore might be used for another signaling application triggered by intracellular or extracellular stimuli.