• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.430-430
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• 2005

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 춘계학술대회
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• pp.171-171
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• 2002

• Biotechnology and Bioprocess Engineering:BBE
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• 제2권2호
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• pp.82-85
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• 1997

The nar promoter as an inducible promoter was characterized for the process development for the gene expression and the protein production under anaerobic condition. The LB medium was selected as a main culture medium showing the enzyme activity of 18,000 units/min/g cell in the flask cultivation. The optimum concentration of nitrate was 1%. Under anaerobic conditions, the gene expression was fully induced in the presence of nitrate.

• 한국자원식물학회지
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• 제3권1호
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• pp.1-30
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• 1990

The traditional plant imprownent methods consisted of pure line selection, cross breeding, heterosis breeding, polyploid breeding, mutati-onbreeding, ect.Biotechmoiogy is divided into gene spliclng , monocle-nal antibodies , protein engineering , agricultural research, and microbiological engineering. Of these , high plants deal with agricultural research, and the importent part of which is tissue culture and celLculture , Tissue .culture and cell culture are again divided into embryoculture, test tube fertilization, anther and pollen culture, somatichybridization , transformation, recombination, recombinant DNA moleculehybrid plasmid, ect For these haploid production, protoplast culture,protoplast fusion, selection and propagation, ect. , the technical sett-lement is needed.

• 한국응용곤충학회:학술대회논문집
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• 한국응용곤충학회 2003년도 추계 학술발표회
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• pp.114-114
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• 2003

• Journal of Plant Biotechnology
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• 제36권4호
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• pp.309-319
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• 2009

Transgenic plant cell cultures for the production of biopharmaceuticals including monoclonal antibodies, recombinant proteins have been regarded as an alternative platform in addition to traditional microbial fermentation and mammalian cell cultures. Plant-made pharmaceuticals (PMPs) have several advantages such as safety, cost-effectiveness, scalability and possibility of complex post-translational modifications. Increasing demand for the quantity and diversity of pharmaceutical proteins may accelerate the industrialization of PMP technology. Up to date, there is no plant-made recombinant protein approved by USFDA (Food and Drug Administration) for human therapeutic uses due to the technological bottlenecks of low expression level and slight differences in glycosylation. Regarding expression levels, it is possible to improve the productivity by using stronger promoter and optimizing culture processes. In terms of glycosylation, humanization has been attempted in many ways to reduce immune responses and to enhance the efficacy as well as stability. In this review article, all these respects of transgenic plant cell cultures were summarized. In addition, we also discuss the general characteristics of plant cell suspension cultures related with bioreactor design and operation to achieve high productivity in large scale which could be a key to successful commercialization of PMPs.

• Journal of Microbiology and Biotechnology
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• 제21권12호
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• pp.1264-1269
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• 2011

Interleukin-2 (IL-2) is a vital cytokine secreted by activated T lymphocytes, and plays an important role in the regulation of cellular functions and immunity of animals. In this study, the recombinant duck IL-2 (rduIL-2) was secretory expressed in Pichia pastoris (P. pastoris). The recombinant P. pastoris strain was cultured in shake flasks and then scaled up in a 5.0-l bioreactor. The result showed that the maximal fresh-cell-weight of 594.1 g/l and the maximal $OD_{600}$ of 408 were achieved in the bioreactor. The rduIL-2 was purified by two steps of purification procedures, and approximately 311 mg of rduIL-2/L fermentation supernatant was obtained. SDS-PAGE showed that the purified rduIL-2 constituted a homogeneous band of ~16 kDa or ~14 kDa corresponding to the glycosylated or non-glycosylated duIL-2 protein in size, respectively. The bioactivity of rduIL-2 was determined by lymphocyte proliferation assay. The result indicated that the rduIL-2 greatly promoted the proliferation of ConA-stimulated lymphocytes in vitro. The P. pastoris expression system described here could provide promising, inexpensive, and large-scale production of the rduIL-2, which lays the foundation for development of novel immunoadjuvants to enhance both the immunity of ducks against various infectious pathogens and vaccine efficacy.

• 한국균학회지
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• 제47권4호
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• pp.359-371
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• 2019

큰느타리버섯에서 철 저장과 관련된 페리틴 단백질의 발현 및 분비를 최적화하기 위해, T-Fer 벡터에 EcoRI 및 HindIII처리를 해 페리틴 유전자를 얻은 후, BamHI으로 처리된 선형의 pPEVPR1b 분비 벡터에 클로닝하여pPEVPR1b-Fer 재조합 벡터를 구축한 다음 Agrobacterium tumefaciens LBA4404 로 도입하였다. Agrobacterium tumefaciens-mediated transformation 방법에 의해 Pleurotus eryngii로 형질전환하고 kanamycin함유된 MCM 배지에서 올바른 형질전환체를 선별하였고, 단백질 발현은 SDS-PAGE 및 항원항체 반응에 의한 western blot으로 확인하였다. 페리틴 단백질의 분비 발현은 batch culture 및 20 L airlift type fermenter에서 배양 시간 및 온도와 같은 배양 조건에 의해 최적화되었다. 페리틴 생산을 위한 배양 조건은 MCM 배지에서 25℃ 및 8 일 배양에 의해 최적화되었다. 페리틴 단백질의 양은 정량적 단백질 분석에 의해 2.4 mg/g mycelium으로 측정되었다. 그러나, PR1b (32 amino acid)의 분비서열은 큰느타리버섯 내부의 peptidase에 의해 정확하게 processing되지 않았지만, 페리틴 단백질은 균사체에서 최대로 전체단백질의 24.7% 발현되었고, 배양액에서는 검출되지 않았다. 철 결합 활성은 7.5% non-denaturing gel에서 Perls' staining에 의해 확인되었으며, 다량체 페리틴(24 subunits)이 P. eryngii 균사체에서 형성되었음을 보여준다. 생물학적 활성 측정을 위하여 페리틴을 함유한 분말을 제조하여 육계의 사료 첨가제로서의 사용 가능성에 대해 시험하였으며, 결과적으로 페리틴은 육계의 성장을 촉진하고 사료 효율 및 생산 지수를 향상시키는것으로 확인되었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.261-264
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• 2002

CHO (Chinese hamster ovary) cells were transfected with plasmids containing both cis-acting HRE (hypoxia response element) and CMV-promoter that controls tissue-type plasminogen activator (t-PA). CHO cells with HRE produced 16.2 fold higher t-PA concentration than CHO cells without HRE. It was noted that hypoxia strongly induced CHO cell apoptosis. which resulted in decrease of cell viability and protein production. In this study. by introducing Bcl-2, anti-apoptotic gene, we tried to recover cell viability and increase the protein production. When batch culture of both control cells without transfection of Bcl-2 and cells transfected with Bcl-2 were performed in the absence of CoCl ι hypoxia mimic condition. the cells with Bcl-2 were effected specific cell growth rates, maximum cell density. Immunoblotting assay showed Bcl-2 was recombinant with HRE dependent t- P A expression cassette, and their expression level was depended on hypoxia. By introducing Bcl-2, both cell viability and maximum cell density could be increased.

• Molecules and Cells
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• 제25권4호
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• pp.494-503
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• 2008

Many therapeutic glycoproteins have been successfully generated in plants. Plants have advantages regarding practical and economic concerns, and safety of protein production over other existing systems. However, plants are not ideal expression systems for the production of biopharmaceutical proteins, due to the fact that they are incapable of the authentic human N-glycosylation process. The majority of therapeutic proteins are glycoproteins which harbor N-glycans, which are often essential for their stability, folding, and biological activity. Thus, several glyco-engineering strategies have emerged for the tailor-making of N-glycosylation in plants, including glycoprotein subcellular targeting, the inhibition of plant specific glycosyltranferases, or the addition of human specific glycosyltransferases. This article focuses on plant N-glycosylation structure, glycosylation variation in plant cell, plant expression system of glycoproteins, and impact of glycosylation on immunological function. Furthermore, plant glyco-engineering techniques currently being developed to overcome the limitations of plant expression systems in the production of therapeutic glycoproteins will be discussed in this review.