• Korean Journal of Optics and Photonics
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• v.35 no.1
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• pp.30-34
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• 2024

We report a purely protein-based platform for green fluorescence by mixing silk protein with green fluorescence protein, and also report its enhancement by the incorporation of TiO₂ nanoparticles. The TiO₂ nanoparticles employed have diameters of 100 and 300 nm, with a significant increase in fluorescence (by a factor of 7.5) observed when introducing 300-nm TiO₂ nanoparticles. Furthermore, an increase in particle distribution density is found to enhance fluorescence amplification. These research findings suggest that protein-based fluorescent films can be enhanced by the characteristics of nanoparticles, opening up new possibilities in the fields of optics and fluorescence applications.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.153-158
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• 2013

To express green fluorescent protein in the cocoon of silkworm, we constructed the fibroin H-chain expression system to produce enhanced green fluorescent protein (EGFP) in the cocoon of transgenic silkworms. The EGFP fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the EGFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven DsRed2 cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,200 eggs of bivoltin silkworms, Baegokjam. We obtained 8 broods. The cocoon displayed strong green fluorescence, proving that the fusion protein was present in the cocoon. Also, the presence of fusion proteins in cocoons was demonstrated by SDS-PAGE and immunoblotting. Accordingly, we suggest that the EGFP fluorescence silk will enable the production of the novel biomaterial based on the transgenic silk.

• Proceedings of the Korean Society of Life Science Conference
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• 2003.05a
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• pp.38-47
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• 2003

처음으로 real time으로 유전자 발현을 visualization할 수 있는 marker를 이용할 수 있게 되었다. 이 maker가 바로 green fluorescent protein(GFP; 녹색형광단백질)이다. GFP는1962년 Shimomura 등에 의하여 해파리인 Aequorea victoria에 존재함이 밝혀졌다(1). 그러나 30년이 지난 1992년이 되어서야 Plasher등에 의하여 GFP의 cDNA가 클로닝 되었고(2) 이후 지금까지 약 10년 동안 GFP는 생명과학분야에서 가장 각광받는 유용한 단백질 중의 하나가 되어 매우 다양한 연구에 응용이 되고 있다. 이렇게 GFP가 생명과학분야에서각광을 받게 된 이유로는 GFP가 모든 외래의 세포에서 형광을 발할 수 있는 활성을 가진 형태로 발현되기 때문이다(3). Chalfie 등은 처음으로 GFP를 대장균과 Caenorhabditis elegans에서 발현시켜 유전자의 발현을 모니터함으로써 GFP를 원핵 및 진핵세포 모두에서 사용할 수 있다는 것을 보고하였다(3), 이러한 발견을 통하여 GFP는 살아있는 세포, 조직 및 생물체에 해를 주지 않고 (non-invasive) 유전자의 발현을 측정하는 marker로서 사용할 수 있게 되어 cell biology, developmental biology, neurobiology 및 cytology 등의 연구분야에 널리 이용되고 있다.

• Annual Conference on Human and Language Technology
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• 2003.10d
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• pp.293-299
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• 2003

인간 게놈 프로젝트가 완료됨에 다라 생체서열과 언어 사이의 대응 관계가 부각되고 있다. 본고에서는 Lewis Carroll의 언어 유희 사례를 컴퓨터생물학의 측면에서 재조명하고, Carroll이 제시한 문제 중에서 간단한 anagram 문제의 해결을 다루고자 한다. 우선 DNA 컴퓨팅의 방법론을 적용한 DNAgram의 개념을 확장하여 plasmid-DNAgram의 개념을 새롭게 도입하였다. 이 개념을 형광단백질에 대한 DNAgram의 개념을 확장하여 plasmid-DNAgram의 개념을 새롭게 도입하였다. 이 개념을 형광단백질에 대한 FRET(fluorescent resonance energy transfer)분석기법의 응용 사례인 cameleon 형광단백질에 대한 FRET 분석기법에 적용함으로써 anagram 문제의 어휘론적, 구문론적, 의미론적, 화용론적 측면에 대응하는 바이오분자 컴퓨팅 방법론을 제안하였다.

• Journal of Industrial Technology
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• v.29 no.B
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• pp.115-119
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• 2009

Green fluorescent protein (GFPuv) was displayed on the surface of ethanol-producing bacteria Zymomonas mobilis using N-terminal domain of ice nucleation protein (INP) as an anchoring motif. To evaluate the ice nucleation protein as plausible anchor motif in Z. mobilis, GFPuv gene was subcloned into Zymomonas expression vector yielding pBBR1MCS-3/pPDC/INPN/GFPuv plasmid., INP-GFPuv fusion protein was expressed in Z. mobilis and its fluorescence was verified by confocal microscopy. The successful display of GFPuv on Zymomonas mobilis suggest that INP anchor motif could be used for future fusion partner in Z. mobilis strain improvement.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.142-146
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• 2013

The fluorescent proteins are generally denatured by heat treatment and thus lose their color. The normal reeling method includes processing by drying and cooking the cocoons near $100^{\circ}C$ before reeling. Therefore, the usual processing method cannot be used for making colored fluorescent silks. To develop a method that is applicable to producing transgenic silk without color loss, we develop reeling methods adequate for a recombinant fluorescence cocoons. It was found that the fluorescence cocoons keep their native color when dried at temperatures lower than $60^{\circ}C$ for 15 h. Also, a new cooking method to soften the fluorescent cocoons was developed: the cocoons were soaked in a solution of 0.2% sodium carbonate ($Na_2CO_3$)/0.1% nonionic surfactant (Triton X100) at $60^{\circ}C$ and then placed under vacuum. The repeated vacuum treatments enabled complete penetration of the solution into the cocoons, and the cocoons were thus homogenously softened and ready for reeling. In this state, the cooked cocoons can be reeled by an automated reeling machine. Our results suggest that drying and cooking of the cocoons at low temperature enables the subsequent reeling of the colored fluorescent silks by an automatic reeling machine without color loss and can produce silks that can be used for making higher value-added silk materials.

• The Science & Technology
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• s.508
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• pp.38-41
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• 2011

• 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.

• Journal of Sericultural and Entomological Science
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• v.52 no.2
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• pp.117-122
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• 2014

On previous studies, we constructed a transgenic silkworm which produces the chimeric silk fused green fluorescent protein (EGFP), but the transgenic silkworm has decreased commercial feasible traits such as convenience of breeding and productivity of silk. In this study, we performed cross fertilization between green fluorescent silk transgenic silkworm and colored cocoon silkworm descents to make the transgenic the transgenic silkworm producing improved fluorescence cocoon. In the result, we found out a bit valuable cross fertilization manners ($female{\times}male$) in respect of silk productivity such as $T59B{\times}Jam26$, $Jam329{\times}T59W$, $T59W{\times}Jam329$, and $T59W{\times}Jam178$. The color-difference of offspring cocoons were measured according to different cross manners using by CIE Lab-based formulae with a X-rite VS450. In the result, the depth of green color of cocoons was a little high at cross manners as $Jam329{\times}T59W$, $T59W{\times}Jam178$. Meanwhile, the depth of yellow clolor of cocoons was remarkable at cross manners as $Jam178{\times}T59W$, $T59W{\times}Jam178$, respectively.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2004.11a
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• pp.64-73
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• 2004

세포 내에서 일어나는 신호 전달 과정은 단백질간의 상호작용을 통해 수행되고 조절된다. 단백질 상호작용 데이터를 활용하여 수행된 연구로는 단백질의 기능을 유추하거나 전체 네트워크 중 다른 지역보다 더 조밀한 상호작용을 추출하여 complex 혹은 pathway를 발견하고 진화 과정을 이해하는 바탕이 되고 있다. 본 연구에서는 신호 전달 경로에 대한 사전 정보 없이 yeast 상호작용 정보와 녹색형광단백질(GFP)을 이용하여 밝혀진 4000여 개의 yeast 단백질 위치 분포 data를 이용하여 신호전달경로를 찾는 방법을 시도했다. 기존 연구에 의해 밝혀진 yeast 내의 단백질 위치 분포 결과를 보면 21개의 category에 대해 각 단백질 상호작용 분포가 다양하게 나타나고, 특정 위치에서 상호작용 빈도수가 현저히 크다는 것을 알 수 있다. 특히 두 단백질이 같은 장소에 있을 경우 상호작용 확률이 높으며, 세포 내 소기관 사이에도 상호작용의 정도가 다양함이 알려져 있다. 따라서 이러한 분포상의 특성을 고려하여 상호작용을 기반으로 하여 세포막 단백질을 출발점으로, 핵에 있는 단백질을 도착점으로 잡고, 그 사이에 존재하는 다양한 가능 경로 중에서 단백질의 위치 정보를 가중치로 사용하여 그 중 최대 가능 경로를 찾도록 구현하였다. 이와 같은 pathway 모델링은 기존에 밝혀진 pathway와의 비교를 통해 알려지지 않은 새로운 경로를 발견하고, 이전에 경로에 참여하지 않은 단백질들을 발견할 수 있고, 이미 알려진 단백질들의 새로운 기능들에 대해서도 추론할 수 있을 것이라 기대한다.