• 식물조직배양학회지
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• 제25권6호
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• pp.501-505
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• 1998

오이의 국내 F1 품종인 조생낙합의 하배축 유래 배발생 현탁배양 세포의 초저온 보존 시스템을 개발하였다. 액체질소 저장 후 캘러스 재생률은 2M DMNSO와 0.4 M sucrose를 혼용 처리하였을 때 캘러스 재생률이 85%로 가장 높았다. 그러나 glycerol 처리구에서는 처리농도에 상관없이 모든 처리구에서 캘러스 재생이 이루어지지 않았다. 또한 고농도의 삼투용액에서 배양세포의 전처리 과정은 필요하지 않았다. 재생된 캘러스를 1 mg/L 2,4-D가 첨가된 MS 배지로 이식하여 배양하였을 때 다수의 체세포배가 발달하였으며, 체세포배를 MS 기본배지로 옮겨 명배양한 결과 다수의 소식물체가 발달하였다.

• Journal of Microbiology and Biotechnology
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• 제8권3호
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• pp.237-244
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• 1998

A study was carried out to elucidate the relation between the production of flavonol glycosides and the change of phenylalanine ammonia-lyase activity in cell suspension cultures of Ginkgo biloba by the unassisted and synergistic effects of various factors. The quercetin production showed a mixed-growth-associated pattern in cell suspension cultures. Fluorescent light and UV radiation increased phenylalanine ammonia-lyase (PAL) activity, and resulted in the increase of the production of quercetin and kaempferol ten- and four-fold, respectively, as compared to that obtained in the normal culture condition. The cell growth of Ginkgo biloba was enhanced .at higher temperatures whereas the quercetin production was at its maximum at low temperatures. Moreover, the quercetin production was increased by temperature change during the culture period. In particular, the quercetin production was at the highest level when the culture temperature was elevated from $10^{\circ}C\;to\;30^{\circ}C$. The addition of phenylalanine as a precursor in the culture medium stimulated an 8-fold increase in the production of quercetin; the addition of naringenin caused a l0-fold increase. The quercetin production was also greatly increased by feeding enzyme cofactors such as 2-ketoglutarate and ascorbic acid in the culture medium, but specific PAL activity was not increased except with phenylalanine feeding. The synergistic effect of UV radiation and naringenin feeding was observed, resulting in the increase of flavonol glycoside production at a rate higher than in any other case investigated.

• Journal of Microbiology and Biotechnology
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• 제13권2호
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• pp.256-262
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• 2003

To enhance the production of heavy chain immunoglobulin G (HC IgG) in the suspension cultures of transgenic tobacco cells (Nicotiana tabacum), the effects of adding various cytokinins (i.e., growth regulators) and organic nitrogen sources to culture media were investigated. Four different cytokinins including kinetin, isopentenyladenine (IPA), 6-benzylaminepurine (BA), and zeatin were tested with or without dichlorophenoxyacetic acid (2,4-D), which is a typical growth regulator supplemented in the standard Murashige and Skoog (MS) medium. The productivity of intracellular HC IgG was increased by 36 and $42\%$, compared to the control, especially when IPA (2 mg/l) or BA (0.2 mg/l) was added to the media in the presence of 2,4-D, respectively. In the study of organic nitrogen sources, addition of each casein hydrolysate and tryptone to the culture media at a final concentration of 0.01 and 1 g/l, respectively. increased the productivity or he IgG as much as 68 and $67\%$, respectively, in comparison with the control, which was is MS medium without supplementation of any organic nitrogen sources. This study shows that the optimization of media composition could offer significant improvements in the production of foreign proteins in the suspension cultures of transgenic plants.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1999년도 춘계학술발표대회:발표논문요지록
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• pp.12-12
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• 1999

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.250-253
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• 2000

We researched effects of growth on initial inoculum size, liquid volume, and medium feeding rate etc. Cell suspension inoculated at low cell concentrations showed a typical growth reduction, whereas root cultures showed an improvement in growth. In this paper, Hairy roots showed high growth rate at 0.4 % inoculum size and 100 mL liquid volume in 250 mL flask cultures.

• KSBB Journal
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• 제8권5호
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• pp.504-507
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• 1993

Control culture에서 embryo 생성률이 6개월 동안 1000embryos/ml에서 500embryo/ml로 감소되었다. 이러한 현상을 극복하기 위한 방법으로 embryo 배양액에서 추출된 세포외 물질의 첨가는 embryo 생생률을 control culture와 비 교 하여 최 대 (2500embryos/ml)까지 증가시켰다. 또한 세포외 단백질이 embryo 수율 향상에 기인하는 것으로 추측된다.

• 한국산림과학회지
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• 제78권2호
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• pp.198-208
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• 1989

버드나무의 잎으로부터 유도(誘導)된 callus의 현탁배양(懸濁培養)은 MS 기본배지(基本培地)에 2,4-D 1.0mg/l와 zeatin 0.1mg/l를 첨가(添加)한 액체배지(液體培地)에서 생장(生長)이 가장 좋았다. 액체배지(液體培地)에서 24일간(日間) 세포(細胞)를 배양(培養)한 후(後) 분리(分離)된 배양액(培養液)은 상치, 벼 그리고 피 종자(種子)의 발아(發芽)를 강(强)하게 억제(抑制)하여 배양중(培養中)에 생리활성(生理活性) 물질(物質)이 형성(形成)되는 것으로 추정(推定)되었다. 배양세포(培養細胞)에서는 pyrogallol, sinapic acid, cinnamic acid, tannic+gallic acid 그리고 P-chlorobenzoic acid 등(等)과 같은 여러 종류(種類)의 phenol 화합물(化合物)이 검출(檢出)되어 이들이 강(强)한 발아(發芽) 억제작용(抑制作用)을 하는 것으로 사료(思料)되었다.

• Journal of Plant Biotechnology
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• 제4권4호
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• pp.143-150
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• 2002

A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권2호
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• pp.162-165
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• 2005

A series of fluorine and hydroxyl containing jasmonate derivatives, which were chemically synthesized in our institute, were investigated for their effects on the biosynthesis and heterogeneity of ginsenosides in suspension cultures of Panax notoginseng cells. Com-pared to the control (without addition of elicitors), $100{\mu}M$ of each of the jasmonate was added on day 4 to the suspension cultures of P. notoginseng cells. It was observed that, jasmonates greatly enhanced the ginsenoside content and the ratio of Rb group to Rg group (i.e. $(Rb_1\;+\;Rd)/(Rg_1\;+\;Re)$ in the P. notoginseng cells. Some of the synthetic jasmonates, such as pentafluoropropyl jasmonate (PFPJA), 2-hydroxyethyl jasmonate (HEJA) and 2-hydroxye-thoxyethyl jasmonate (HEEJA), could promote the ginsenoside content to $2.55\;\pm\;0.11,\;3.65\;\pm\;0.13\;and\;2.94\;\pm\;0.06$mg/100 mg DW, respectively, compared to that of $0.64\;\pm\;0.06$mg/100 mg DW for the control and $2.17\;\pm\;0.04$ mg/100 mg DW by the commercially available methyl jasmonate (MJA); and they could change the respective Rb:Rg ratio to $1.60\;\pm\;0.04,\;1.87\;\pm\;0.01\;and\;1.56\;\pm\;0.05$, compared to that of $0.47\;\pm\;0.01$ for the control and $1.42\;\pm\;0.06$ by MJA. The results suggest that suitable esterification of MJA with fluorine or hydroxyl group could in-crease the elicitation activity to induce plant secondary metabolism. The information obtained from this study is useful for hyper-production of heterogeneous products by plant cell cultures.

• KSBB Journal
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• 제15권5호
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• pp.428-433
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• 2000

희수 (Camptotheca acuminata) 현닥세포배양에서 최근 항암 제로서 관심을 팔고있는 camptothecin을 얻기 위하여 연구를 수행하였다. 현탁세포배양에서 exponential phase의 최고 생장 속도는 $0.269day^{-1}$이었으며 배가시간이 2.58일이었다. Camptothecin 은 배양초기에 생성되지 않고 exponential phase 말기부터 서서히 생성되어 stationary phase 초기에 최고를 나타내고 이후 급격히 감소하였다. Camptothecin의 생산을 촉진하기 위하여 elicitor를 사F용하였다. 여러 elicitor 중 jasmonic acid와 cellulase 가 camptothecin의 생성을 증가시켰다, Elicitor는 접종과 동시 에 투여 했을 때 가장 효과가 좋았으며, jasmonic acid와 cellulase 의 combination으로 효과가 더욱 증진했는데 대조구에 비하여 약 20배 정도 증가한 $20.42{\times}10^{-3}mg/\ell$ 의 camptothecin이 생 성되었다. 이것은 생산량을 증가시킨 것뿐만 아니라 생산기 간도 약 1/4로 단축시키는 결과를 얻을 수 있었다.