• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.87-90
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• 2000

• The Microorganisms and Industry
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• v.15 no.2
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• pp.46-49
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• 1989

단백질의 아미노산 서열에 변화를 줄려고 할 때 대상단백질의 3차구조에 대한 충분한 정보가 있고 시험해 보고자 하는 가설이 확실할 때에는 특정잔기를 다른 특정잔기로 치환시켜 효능을 전환시키는 것이 효과적이겠고, 3차구조가 안밝혀져 있거나 어떻게 치환해야 할지 모를 경우에는 무작위 변이유도법과 세포에 의한 형질선별법이 바람직하다 하겠다. 이러한 관점에서 볼 때 유전자 재조합기법을 통한 신물질창출을 성공적으로 수행하기 위해서는 유전자 조작기술과 단백질 분석기술은 물론 단백질생화학, 분자유전학, 미생물생리학 등에 대한 충분한 이해를 바탕으로하여 여러 분야에서 협동적으로 연구되어야 하겠다.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.05a
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• pp.101-103
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• 2001

가식성 및 생분해성 필름은 식품과 외부와의 물질이동을 차단하고, 영양성분, 향기성분, 항 산화제 등의 carrier 역할을 한다. 기존의 플라스틱 고분자 필름은 생분해성 필름에 비해 가격 경쟁력 및 물리적 성질이 우수하기는 하지만, 생분해성 필름은 단백질과 다당류 및 지질을 주원료로 하여 이들의 단독 또는 조합에 의해 만들어지고, 생물학적 분해가 용이하고 식용 가능하여 포장재로 인한 환경오염을 줄일 수 있다는 것이 생분해성 필름의 가장 큰 장점이다. (중략)

• Journal of Life Science
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• v.20 no.12
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• pp.1743-1749
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• 2010

A research group demonstrated that the 37 kDA protein of Edwardsiella tarda, a causing causative agent of edwardsiellosis in fish, exhibited high antigenicity in Japanese flounder. The research group also showed that the N-terminus amino acid sequences of the 37 kDa protein were mapped to the N-terminus of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Using degenerated primer sets based on the known N-terminus sequence, the corresponding E. tarda DNA was amplified and cloned. The nucleotide sequences of the cloned gene revealed high homology with a bacterial gene for GAPDH, as we was expected. The amino acid sequence of E. tarda GAPDH (etGAPDH) revealed a <70% similarity with GAPDH proteins in other Enterobacteriaceae. With the application of artificial protein overexpression system in Escherichia coli, the recombinant etGAPDH (rGAPDH) was produced and purified. In this study, Using the purified rGAPDH, the etGAPDH specific polyclonal antibody has been was generated using the purified rGAPDHin this study. The immunoblotting analyses demonstrated that the location of the GAPDH protein is located with the association of is associated with the envelops of E. tarda. The rGAPDH was administrated into Japanese flounder via IP route for evaluation of the protective ability. Although the specific antibody titer against etGAPDH was increased about 3-fold after 4 weeks post-vaccination, the survival rates of vaccinated Japanese flounder and the control group with wild type E. tarda was were 12.5% and 0%, respectively. Our results indicated that rGAPDH is immunoreactive antigen but that it will not generate protective immunity in Japanese flounder.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.711-714
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• 2001

The production of the recombinant Plasmodium vivax merozoite surface protein (PvMSP) has been investigated in the recombinant E.coli system. Experimental optimization of the culture conditions, such as the effect of initial pH, and operating temperature has been tried on the growth of recombinant E.coli and on the overproduction of the target foreign protein.

• Environmental Analysis Health and Toxicology
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• v.19 no.3
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• pp.295-305
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• 2004

본 연구에서는 유전자 재조합 발광 박테리아를 이용하여 농약에 대한 박테리아의 스트레스 반응과 세포 독성을 분석하였다. 15종류의 농약에 대하여 유전자 손상, 생물막 손상, 산화적 손상 및 단백질 손상을 측정할 수 있는 발광 박테리아와 독성 유무로 인한 세포 독성을 측정할 수 있는 발광 박테리아, 5종을 이용하여 스트레스 반응을 분류하고 세포 독성 정토를 분석하였다. 그 결과, 농약의 화학적 구조가 박테리아의 스트레스 반응에 영향을 미치며, 산화과정이 진행 됨에 따라 독성의 작용 기작이 변하는 것을 확인 할 수 있었다. 이와 같은, 유전자 재조합 발광 박테리아를 이용한 생물체내의 독성 메커니즘에 대한 분석은 생태계 유해물질들에 의한 독성을 분석하고 예상하기 위해 적용될 수 있을 것이다.

• Journal of Life Science
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• v.28 no.3
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• pp.275-283
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• 2018

Human fibroblast growth factor (FGF) has the potential to be a commercially important therapeutic or cosmeceutical agent due to its ability to generate tissue and heal wounds. Granting permeability into skin tissues increases the therapeutic effects of FGF. Thus, several researchers have attempted the fusion of FGF conjugates with protein transduction domains (PTDs) to investigate the transduction ability and therapeutic effects of FGF. Less is known, however, about whether the location of PTD fused to the N- or C-terminus of FGF proteins has a significant impact on the folding and stability in Escherichia coli, and eventually, on transduction. Here, we report cloning of human basic fibroblast growth factor (FGF2) as a control and FGF2 with PTD fused to the N- or C-terminal ends of FGF proteins by an overlap extension PCR. We performed expression, verified expression properties of recombinant FGF2 without or with PTD fused to the N-terminus and the C-terminus, and investigated transduction ability into tissue by treating the dorsal skin of mice subjects. As a result, FGF2 and FGF2-PTD (fused to C-terminus) fusion protein were expressed as soluble forms suitable for straight-forward purification, unlike insoluble PTD-FGF2 (fused to N-terminus), but only FGF2-PTD fusion protein could transduce into the dorsal skin tissue of the mice subjects. Our results suggest that FGF2 with PTD fused to the C-terminus is more efficient than other options in terms of expression, purification, and delivery into skin tissue, as it does not require labor-intensive, costly, and time-consuming methods.

• KSBB Journal
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• v.15 no.2
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• pp.174-180
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• 2000

Pichia pastoris and Hansenula polymorpha, the methylotrophic yeasts have been widely used as a host for the production of e eudaryotic proteins due to the advantages related to their inherited characters. This paper describes the method to enhance t the productivity of recombinant proteins by P. pastoris and H. po$\psi$morpha. In the production of recombinant proteins using a f fed batch fermentation system, the effects of specific growth rate on the specific expression rate of re$\infty$mbinant proteins w were studied. In both species, the expression system of recombinant proteins using the fed batch fermentation was optimezed.

• Journal of Life Science
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• v.22 no.8
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• pp.1024-1033
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• 2012

A gene encoding a novel geranylgeranyl pyrophosphate (GGPP) synthase from Kocuria gwangalliensis has been cloned and expressed in Escherichia coli. The deduced amino acid sequence showed 59.6% identity with a putative GGPP synthase (CrtE) from K. rhizophila. An expression plasmid containing the crtE gene was constructed, and E. coli cells containing this plasmid produced a recombinant protein with a theoretical molecular mass of 41 kDa, corresponding to the molecular weight of GGPP synthase. Due to the lack of crtE, crtB, and crtI in E. coli, the biosynthesis of lycopene was only obtained when the plasmid pCcrtE was co-transformed into E. coli expressing the pRScrtBI-carrying carotenoid biosynthesis crtB and crtI genes, which were sub-cloned from Paracoccus haeundaensis. The biochemical studies on the expressed proteins were performed via HPLC. The results obtained from this study will provide a wider base of knowledge regarding the primary structure of CrtE cloned from K. gwangalliensis at the molecular level.

• Journal of Food Hygiene and Safety
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• v.18 no.1
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• pp.25-29
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• 2003

The p60 protein of Listeria spp. is a Listeria-Genus-specific, major extra-cellular protein, which is used as an indicator protein for the detection of these bacteria from contaminated foods. In this study, p60 protein were recombinantly produced in E. coli and were purified using amylose resin based column chromatography. Purified recombinant-p6O was used to generate monoclonal antibody against native p60. Antibody from hybridoma cell line, 1H4, specificically reacted with native p60 protein isolated from pathogenic Listeria spp. such as L. monocytogenes, L. ivanovii, L. welshimeri II, but did not or relatively weakly reacted with non-pathogenic Listeia species, L. innocua or other bacterial proteins. Antibody from 1H4 was produced using ascites fluid method and it may be useful to develop the Listeria-detection kits based on immunological method.