• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.239-239
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• 2004

본 연구에서는 효과적인 유전자를 발현시킬 수 있는 retrovirus vector system을 구축하기 위하여 네 가지 promoter를 비교하였다. 일반적으로 사용되고 있는 RSV (Rous sarcoma virus), UbC (Ubiquitin), β-actin, CMV(cytomegalovirus) promoter 하에 GFP (green fluorescent protein)를 표지유전자로 사용하였다. 또한 WPRE (Woodchuck hepatitis virus Posttranscriptional Regulatory Element) 서열을 도입하여 각각의 promoter 하에서 GFP 유전자의 발현 증가 여부를 검정하였다. (중략)

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.823-826
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• 2001

In the initial process of photosynthesis, a biological electron transfer system, photoelectric conversion occurs and then long-range electron transfer takes place very efficiently in one direction through the biomolecules. The metal/insulator/metal structured device consisting of GFP, viologen, cytochrome c hetero-thin film was presented based on the biomimesis. GFP, viologen, and cytochrome c was used as an electron sensitizer, a mediator, and an electron acceptor. Cytochrome c molecules and viologen molecules were deposited by Langmuir-Blodgett (LB) technique, and GFP molecules were adsorbed by self-assembly method (SAM). Surface morphology of hetero-thin film was analyzed by scanning tunneling microscopy (STM). Local photoswitching effects of a proposed photodiode were verified by current-voltage measurements using hybrid STM/I-V measurement system.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2004년도 제21차 정기총회 및 학술발표회
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• pp.19-20
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• 2004

본 연구는 1세포기 닭 수정란에서 외래 표지유전자(EGFP)와 리포좀(liposome)을 사용하여 외래유전자의 핵 전이의 효율성을 검증하고자 하였다. 실험은 리포좀과 혼합된 표지유전자와 naked 유전자 두 그룹으로 나누고, 유전자 미세 주입방법을 이용하여 배반엽 단계(stage X)와 1세포기 수정란의 세포질에 미세 주입하고 지속적으로 배양하면서 GFP의 발현 양상들을 관찰하였다. 실험결과, 배반엽 단계와 1세포기 수정란 모두에서 리포좀과 외래 유전자의 혼합물을 미세 주입한 경우 일주일 정도 지속적으로 GFP가 발현되었으나, 외래 유전자만을 주입한 경우 GFP의 발현이 관찰되지 않았다. 본 연구결과는 리포좀이 효율적으로 외래 유전자를 닭 수정란의 핵으로 이동시킴을 보여주고 있다.

• 대한수학회논문집
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• 제25권1호
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• pp.139-147
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• 2010

A generalized nondifferentiable fractional optimization problem (GFP), which consists of a maximum objective function defined by finite fractional functions with differentiable functions and support functions, and a constraint set defined by differentiable functions, is considered. Recently, Kim et al. [Journal of Optimization Theory and Applications 129 (2006), no. 1, 131-146] proved optimality theorems and duality theorems for a nondifferentiable multiobjective fractional programming problem (MFP), which consists of a vector-valued function whose components are fractional functions with differentiable functions and support functions, and a constraint set defined by differentiable functions. In fact if $\overline{x}$ is a solution of (GFP), then $\overline{x}$ is a weakly efficient solution of (MFP), but the converse may not be true. So, it seems to be not trivial that we apply the approach of Kim et al. to (GFP). However, modifying their approach, we obtain optimality conditions and duality results for (GFP).

• 한국해양바이오학회지
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• 제5권4호
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• pp.26-32
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• 2011

Green fluorescent protein (GFP), a very important biological agent that involves shifting the color of bioluminescence from blue to green in luminous coelenterates and to increase the quantum yield of light emission. GFP discovered in medusa, Aequorea victoria is a key factor of various biotechnological and cell biological applications. Beside these applications, GFP of A. victoria is generally stable, which does not require co-factors for activity and can be functionally expressed in different bacterial species. This property of GFPs from A. victoria permits them to be a unique tool to monitor gene expression and protein localization in different organisms. The present review brings out the past milestones and future perspectives on GFPs, with an elaborative reviewing on its applications.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.188-188
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• 2004

근래에 많은 형질전환 실험동물이 생산되어 인간을 대체하여 전임상실험에 사용되고 있으며, 그 요구는 점차 증가되고 있다. 그러나 지금까지 생산된 것은 소형 실험동물에 국한되고 있어, 사람과의 그 크기와 기능에 거리가 있었던 것이 사실이다. 본 실험은 중형 실험동물이자 사람과 여러 기관 및 조직이 유사하다고 알려진 돼지로부터 난자를 채취하여 난자의 핵을 GFP가 발현된 세포의 핵으로 치환하여 태어난 산자를 재복제하여 생산된 돼지 신생자를 이용하여 각 기관 및 조직에서 GFP의 발현을 RT-PCR, Northern blot 및 immunocytochemistry 방법으로 분석하였다. (중략)

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.176.2-176.2
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• 2006

• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.358-363
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• 2008

The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.797-804
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• 2000

The human immunodeficiency virus type 1 (HIV-1) Tat is one of the viral gene products essential for HIV replication. The exogenous Tat protein is transduced through the plasma membrane and then accumulated in a cell. The basic domain of the Tat protein, which is rich in arginine and lysine residues and called the protein transduction domain (PTD), has been identified to be responsible for this transduction activity. To better understand the nature of the transduction mediated by this highly basic domain of HIV-1 Tat, the Green Fluorescent Protein (GFP) was expressed and purified as a fusion protein with a peptide derived from the HIV-1 Tat basic domain in Escherichia coli. The transduction of Tat-GFP into mammalian cells was then determined by a Western blot analysis and fluorescence microscopy. The cells treated with Tat-GFP exhibited dose- and time-dependent increases in their intracellular level of the protein. the effective transduction of denatured Tat-GFP into both the nucleus and the cytoplasm of mammalian cells was also demonstrated, thereby indicating that the unfolding of the transduced protein is required for efficient transduction. Accordingly, the availability of recombinant Tat-GFP can facilitate the simple and specific identification of the protein transduction mediated by the HIV-1 Tat basic domain in living cells either by fluorescence microscopy or by a fluorescence-activated cell sorter analysis.

• The Plant Pathology Journal
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• 제27권4호
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• pp.310-314
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• 2011

Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.