• 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.

• Journal of Environmental Science International
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• v.6 no.6
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• pp.595-604
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• 1997

To investigate characteristics of biogeochemical environment of the Korea Deep Ocean Study(KODOSI area in the northeast Pacific Ocean, we preferentially measured Inorganic nutrients and fluorescent organic matters. Typically. the permanent thermocline was well developed at the depth of 200~1000m In the study area. Nitrate. phosphate and silicate were low In the surface mixed layer and Increased with depth. N/P and N/Si showed 15 and 0.2 respectively In the deeper layer. Two fluorophores, biomacromolecule(protein-like) and geomacromolecule (humid-like) , were observed by three dimensional fluorescence excltatlon/ emission spectra matrix. Biomacromolecule(maximum fluorescence at $Ex_{280m}/Em_{330nm}$) ranged from 41.9 to 147.0 TU with its maximum In the surface mixed layer and minimum in deeper water, This is a same trend that has been reported for DOC in the equatorial Pacific. This suggests that biomacromolecule might be labile and converted to refractory humic substance after bacterial degradation In the deeper layer. On the contrary, geomacromolecule(maximum fluorescence at $Ex_{330m}/Em_{430m}$), ranged from 7.6 to 46.5 QSU, showed minimum in the surface nixed layer(euphotic zone) Implying photodegradation and then increased with depth at all stations. In the characteristics of vertical profiles, the relationship between biomacromolecule and geomacromolecule showed negative correlation. Such trend can be attributed to biochemical regeneration or formation of fluorescent materials accompanying oxidation and rennnerallzation of settling organic matter.

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2001.11a
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• pp.142-142
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• 2001

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2002.11a
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• pp.154-154
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• 2002

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2002.10a
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• pp.106-106
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• 2002

No Abstract, See Full Text

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.04a
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• pp.101-101
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• 2003

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.100-100
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• 2002

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2000.10a
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• pp.53.1-53
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• 2000

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.96-103
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• 2010

Oligosaccharyltransferase (OTase) catalyzes the transfer of a lipid-linked oligosaccharide (LLO) to the nascent polypeptide. Most eukaryotes have an OTase composed of a multisubunit protein complex. However, the kinetoplastid Leishmania major and the bacterium Campylobacter jejuni have only a single subunit for OTase activity, Stt3p and PglB, respectively. In this study, a new in vitro assay for OTase was developed by using a fluorescent peptide containing N-glycosylation sequon, Asn-Xaa-Thr/Ser, where Xaa can be any amino acid residue except Pro. L. major Stt3p and C. jejuni PglB as a model OTase enzyme demonstrated the formation of glycopeptides from a fluorescent peptide through OTase activities. For separation and measurement of the glycopeptides produced by the OTases, Tricine-SDS-PAGE, a lectin column and fluorospectrophotometer, and HPLC were applied. Comparison of these assay methods for analyzing a fluorescent glycopeptide showed HPLC analysis is the best method for separation of glycopeptides and nonglycosylated peptides as well as for quantify the peptides than other methods.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1630-1636
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• 2010

During the fermentation process of Saccharomyces cerevisiae, yeast cells must rapidly respond to a wide variety of external stresses in order to survive the constantly changing environment, including ethanol stress. The accumulation of ethanol can severely inhibit cell growth activity and productivity. Thus, the response to changing ethanol concentrations is one of the most important stress reactions in S. cerevisiae and worthy of thorough investigation. Therefore, this study examined the relationship between ethanol tolerance in S. cerevisiae and a unique protein called alcohol sensitive RING/PHD finger 1 protein (Asr1p). A real-time PCR showed that upon exposure to 8% ethanol, the expression of Asr1 was continuously enhanced, reaching a peak 2 h after stimulation. This result was confirmed by monitoring the fluorescence levels using a strain with a green fluorescent protein tagged to the C-terminal of Asr1p. The fluorescent microscopy also revealed a change in the subcellular localization before and after stimulation. Furthermore, the disruption of the Asr1 gene resulted in hypersensitivity on the medium containing ethanol, when compared with the wild-type strain. Thus, when taken together, the present results suggest that Asr1 is involved in the response to ethanol stress in the yeast S. cerevisiae.