• The Korean Journal of Zoology
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• v.38 no.1
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• pp.106-114
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• 1995

배추흰나비의 조직과 발생시기에 따른 발생 특이 큐티클단백질의 질적 분포를 조사하기 위해 전기영동법, western blotting 및 autoradiography 법을 사용하였다. SDS-PAGE에서 유충기에는 4개, 용기에는 3개의 단백질이 발생 특이 큐티클단백질로 확인되었다. 전기영동적 이동도로 보아 유충기의 62 Kd 큐티클단백질은 지방체 표피, 혈림프에 모두 분포하며. 22 Kd, 16 Kd 단백질은 지방체와 표피, 그리고 19 Kd 단백질은 지방체에서 확인되고 있으며, 용기의 37 Kd 단백질은 표피에서. 28 Kd, 27 Kd 단백질은 표피와 지방체에 분포하는 양상을 보이고 있다 면역학적 방법으로 발생 특이 큐티클단백질의 동질성을 조사한 결과 용기의 27 Kd 단백질은 표피와 지방체에서 반응을 나타내며, 유충기의 22 Kd 단백질은 단지 표피에서만 미약하게 이의 동질성을 보여주고 있다 즉, 27 Kd 단백질은 지방체 22 Kd 단백질은 표피에서 기원하고 있어 발생 특이 큐티클단백질의 합성부위는 한 조직에서 기원하는 것이 아니라 각기 상이한 조직으로 부터 생성되는 것으로 사료된다.

• Journal of Sericultural and Entomological Science
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• v.37 no.2
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• pp.176-180
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• 1995

To characterize expression and formation of three type crystal proteins in transformant, Bacillus thruingiensis PT0529 was analysed by transmission electron microscope and SDS-PAGE according to growth. The results showed that the introduced crystal protein genes, rcyIVD and cytA, were well expressed at earlier stage than resident crystal proteins were also expressed with their own morphology. However, resident crystal protein of B. thuringiensis PT0529 was smaller than that of wild type B. thuringiensis NT0423, suggesting that resident crystal protein production was interfered with introduced two type crystal protein genes.

• Korean journal of applied entomology
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• v.36 no.4
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• pp.341-350
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• 1997

We have now constructed a novel recombinant baculovirus producing fusion protein with Autographa californica nuclear polyhedrosis virus (AcNPV) polyhedrin and Bacillus thuringiensis(Bt) cryIA(c) crystal protein. The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The N-terminal of cryIA(c) gene of Bt subsp. kurstaki HD-73 was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front of intact polyhedrin gene or by insertion into the HindIII site in polyhedrin gene. The recombinant baculoviruses were named as BtrusI or BtrusII, respectively. Although single transcript from the fusion protein gene was apparently observed. BtrusI was produced the two proteins, 92 kDa fusion protein and only polyhedrin. In addition, fusion protein produced by BtrusI did not form polyhedra. Interestingly, however, the cells infected with BtrusII did not show a 33 kDa polyhedrin band as a cells infected with BtrusI. Cells infected with BtrusII were only produced fusion protein, but the polyhedra formed by fusion protein was not observed. To determine the insecticidal toxicity of fusion protein, therefore, Sf9 cells infected with BtrusI were inoculated to Bombyx mori larvae. Sf9 cells infected with BtrusI that expressed the fusion protein caused larval mortality although the insecticidal toxicity was low. In conclusion, our results clearly demonstrated that the fusion protein with polyhedrin and Bt cryIA(c) crystal protein have a insecticida toxicity.

• Applied Microscopy
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• v.18 no.2
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• pp.47-58
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• 1988

두가지 쌀 품종(品種) 종자(種子)(S-201, IR-8)의 호분층을 주사(走査) 및 투과(透過) 전자현미경(電子顯微鏡)으로 관찰(觀察)하면 구형(球形)과 결정형(結晶形)의 단백질체(蛋白質體)가 있는 것으로 밝혀졌으며, 구형(球形) 단백질체(蛋白質體)는 세포질내(細胞質內)에 존재(存在)하고 결정형(結晶形) 단백질체(蛋白質體)는 액포내(液胞內)에서 관찰(觀察)되었다. 가상적(假想的)인 한발 스트레스로서 열자극 처리(處理)($40^{\circ}C$ 에서 4시간)는 호분층(糊粉層)내에 단백질체(蛋白質體) 수에서 정상적(正常的)인 대조구보다 $40{\sim}50%$ 감소(減少)되었다. 또한 호분층(糊粉層)에서 단백질체(蛋白質體) 수의 감소(減少)는 IR-8품종(品種)보다 S-201품종(品種)에서 더욱 현저(顯著)하게 관찰(觀察)되었다. 열자극 처리(處理)로서 단백질체(蛋白質體) 수의 감소(減少)는 S-201품종(品種)에서 결정형(結晶形) 단백질체(蛋白質體)를 둘러싸고 있는 tonoplast membrane의 손상(損傷)으로 밝혀졌으며, 이런 손상(損傷)은 IR-8품종(品種)에는 좀 덜하다는 것으로 나타났다. 즉 이것은 S-201품종(品種)이 IR-8품종(品種)보다 열 자극(刺戟)(한발 스트레스)에 더욱 민감(敏感)하다는 것으로 사료된다. 두 품종(品種)의 종자(種子) 내배유(內胚乳)에 녹말(綠末)이 가득찬 주사(走査) 전자현미경(電子顯微鏡) 사진(寫眞)은 녹말(綠末)이 내배유(內胚乳)의 중앙(中央)으로부터 사출(査出)되는 hexagonal rods로 구성(構成)되어 있다는 것을 나타낸다. 이러한 hexagonal rods는 rods부터 쉽게 분쇄될 수 있는 triangular sectors로 구성(構成)되어 있으며, 이 sectors들의 각 내부(內部)는 $2{\sim}8$개의 단위(單位)로 구성(構成)되어 있는 커다란 compound starch grains들이 들어 있다. 이것은 쌀 내배유세포(內胚乳細胞)에서 compound starch drains들의 매우 다양한 크기를 설명(說明)하고 있다.

• Proceedings of The Korean Society of Health Promotion Conference
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• 2005.11a
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• pp.234-243
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• 2005

상당히 성공적인 사람병원체인 결핵균을 Robert Koch가 발견한 이래, 결핵 방제의 상당한 진보에도 불구하고, 결핵의 발생은 현재까지 계속되어오고 있다(Tiruviluamala and Reichman 2002). 그러므로, 완전한 민감성과 특이성을 갖는 신속한 진단방법이 결핵과 결핵의 잠복감염을 진단하는데 요구되어지며(Tiruviluamala and Reichman 2002), 계속적으로 결핵균과 BCG의 유전체와 단백질체 연구를 통하여 결핵 방제수단이 발전되어질 것이다 (Jungblut et al. 1999). 본 연구는 PPD가 결핵을 진단하는 특이한 항원만으로 정확하게 조화를 이루고 있지 않다는 것을 보여주었으며, 그것의 역가는 결론적으로 생물학적으로 평가되어져야 하지만, PPD의 제조는 단백질체 분석 기법에 의해 분석되고 표준화될 수 있음을 보여주었다. 이를 통하여 결핵 피내진단의 일괄된 결과를 위해 PPD 사용 전에 이와같은 단백질체 분석기법을 적용할 수 있을 것으로 사료되며, 본 연구에서 밝혀진 특이 항원은 새로운 피내진단항원으로 적용될 수 있을 것으로 사료된다.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.173-175
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• 2004

단백질체(Proteome)이란 말은 어원적으로 단백질(protein)에 전체란 뜻을 가진 어미(-body, -some)가 연결된 합성어로 주어진 순간에 세포나 조직이 발현하는 모든 단백질의 총체를 의미하고 이를 연구하는 학문은 Proteomics라 일컫는다. 2001년 2월 International Human Genome Project에 의해 human genome sequence가 밝혀짐으로써 유전체 연구는 일단락 완성되었지만, 염기서열만 가지고는 이 유전자 산물의 기능을 알 수 없었고, 이것이 전사되고 최종적으로 완벽한 모양이 갖추어진 단백질을 분석해야만 그 기능을 알 수 있었다. (중략)

• Korean journal of applied entomology
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• v.38 no.2
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• pp.139-144
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• 1999

We have now constructed a novel recombinant baculovirus producing fusion protein with Autogrqha c.uliforrzica nuclear polyhedrosis virus (AcNPV) polyhedrin and green fluorescent protein (GFP). The fusion protein expressed by the recombinant baculovirus in insect cells was characterized. The GFP gene was introduced under the control of polyhedrin gene promoter of AcNPV, by fusion in the front or back of intact polyhedrin gene. The recombinant baculoviruses were named as Ac-GFPPOL or Ac-POLGFP. respectively. As expected, the 56 kDa fusion protein was expressed in the recombinant virus-infected cells. Interestingly. however, the fluorescence of GFP in the cells infected with Ac- POLGFP was only detected within the nuclei. and that was observed as polyhedra-like granular particles. In the microscopy of cells infected with Ac-GFPPOL, furthermore, GFP was detected in both cytoplasm and nuclei although most of GFP were present within the nuclei. However, fusion protein produced by recombinant virus did not form polyhedra although the fusion protein was fused with polyhedrin and GFP. It is suggested that difference of GFP location in the infected cells appear to be involved in the region of polyhedrin in the fusion protein, and the polyhedrin in the fusion protein might be responsible for the polyhedra-like granular particles present within nuclei.

• Korean journal of applied entomology
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• v.27 no.4
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• pp.211-218
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• 1988

Polyhedral proteins and the endogenous alkaline protease associated with larval-derived polyhedra of nuclear viruses isolated from Spodoptera litura, Bombyx mori, and Hyphantria cunea were investigated. Polyhedral proteins prepared under alkaline protease heat-inactivated condition were separated as one band with 31Kd in S. litura a H. cunea NpV and 30Kd in B. mori NPV by the SDS-polyacrylamide gel electroptoresis. Whereas polyhedral proteins without heat-inactivation were degraded into smaller polypeptides with a certain pattern in alkaline solution. The results of double-immunodiffusion and western blot analysis with antisera against polyhedral proteins indicated that those three polyhedral proteins had common antigenic determinants and the degradation of polyhedral proteins by alkaline protease could be confirmed.

• Journal of Plant Biotechnology
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• v.48 no.3
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• pp.165-172
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• 2021

The solubilization of isolated proteins into the adequate buffer containing of surfactants is primary step for proteomic analysis. Particularly, sodium dodecyl sulfate (SDS) is the most widely used surfactant, however, it is not compatible with mass spectrometry (MS). Therefore, it must be removed prior to MS analysis through rigorous washing, which eventually results in inevitable protein loss. Recently, photocleavable surfactant, 4-hexylphenylazosulfonate (Azo), was reported which can be easily degraded by UV irradiation and is compatible with MS during proteomic approach using animal tissues. In this study, we employed comparative label-free proteomic analysis for evaluating the solubilization efficacies of the Azo and SDS surfactants using rice leave proteins. This approach led to identification of 3,365 proteins of which 682 proteins were determined as significantly modulated. Further, according to the subcellular localization prediction in SDS and Azo, proteins localized in the chloroplast were the major organelle accounting for 64% of the total organelle in the SDS sample, while only 37.5% of organelle proteins solubilized in the Azo were predicted to be localized in chloroplast. Taken together, this study validates the efficient solubilization of total protein isolated from plant material for bottom-up proteomics. Azo surfactant is suitable as substitute of SDS and promising for bottom-up proteomics as it facilitates robust protein extraction, rapid washing step during enzymatic digestion, and MS analysis.

• Journal of Plant Biotechnology
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• v.43 no.2
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• pp.223-230
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• 2016

Ascorbic acid (AsA) is a strong antioxidant/reducing agent that can be converted to dehydroascorbate (DHA) by oxidation in plants. DHA, a very short-lived chemical, is recycled to AsA by dehydroascorbate reductase (DHAR). Previously, DHAR cDNA was isolated from the hairy roots of the sesame plant, and DHAR-overexpressing transgenic potato plants were generated under the control of the CaMV35S promoter (CaMV35S::DHAR). An increase in transgene expression and ascorbate levels were observed in the transgenic plants. In the present study, proteomic analysis revealed that transgenic plants not only accumulated DHAR in their cells, but also induced several other antioxidant enzyme-related proteins during plant growth. These results suggest that DHAR is important for stress tolerance via induction of antioxidant proteins, and could improve stress tolerance in transgenic potato plants.