• 한국동물학회:학술대회논문집
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• 한국동물학회 2001년도 한국생물과학협회 학술발표대회
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• pp.218.3-219
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• 2001

No Abstract, See Full Text

• 한국동물학회:학술대회논문집
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• 한국동물학회 2003년도 한국생물과학협회 학술발표대회
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• pp.193.4-193
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• 2003

No Abstract, See Full Text

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.154-159
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• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

• Journal of Ginseng Research
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• 제24권3호
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• pp.128-133
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• 2000

인삼(Panaxginseng)의 물추출물로부터 분리한 분자량 10 kDa 이상의 분획에서 강한 비장세포 분열증식 효과를 볼 수 있었다. 또한 이 분획은 LPS와 Con A에 의해서 유도된 B-cell과 T-cell의 증식효과를 66%와 49%씩 더 증가시켰다. 이 분획을 80% ammonium sulfate 침전법을 실시해서 조 단백질 분획을 얻었으며 이를 다시 DEAE Sepharose CL-6B column chromatography를 수행해서 단백질 분획들을 분리하였다. 대부분의 단백질은NaCl gradien에 의해서 용출 되었고 0~1 M NaCl gradient에 의해서 분리된 4종류의 당단백질 분획에서 비장세포 증식효과와 대식세포의 NO 생산촉진 효과가 나타났다. 특히 단백질 35.9 %,중성당 49.4%, 산성당 12.5%로 구성되어있는 F-2 분획에서는 LPS수준의 높은 면역세포 증식효과를 보였다. 그러나 F-2를 protease로 처리한 후의 비장세포 증식 효과는 대조군과 대비해서 29.1%가 낮아졌다. 이상의 결과로부터 인삼의 물추출물에서 분리된 F-2분획의 면역세포 활성효과에는 단백질 부분이 중요한 역할을 한다고 생각된다.

• Journal of Plant Biotechnology
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• 제4권3호
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• pp.117-122
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• 2002

Cotton Glutathione S-Transferase(GST: EC 2.5.1.18) was cloned and Gh-5 cDNA was overexpressed in tobacco (Nicotiana tabacum) plants. The transformation of cotton GST in tobacco plant was confirmed by northern blot analysis. Type I and Type II transcript patterns were identified in Gh-5 transgenic tobacco plants. Type I transcripts was only discussed in this paper. Glutathione and 1-chloro-2,4-dinitrobenzene (CDNB) were used as the substrates, and the activity of GST in the type I transgenic plants was about 2.5-fold higher than the non-expressers and wild type tobacco plants. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type I transgenic plants produced functional GST in the cells. Type I showed higher GST specific activity than Type II in the transgenic plants. Control and transgenic seedlings were grown in the growth chamber and under the light at 15$^{\circ}C$, and the effects of cotton GST in the seedlings was evaluated. The growth rate of Gh-5 overexpressors was better than the control and non-transgenic tobacco plants. Salinity tolerance was also analyzed on the seeds of transgenic plants. Seeds of Gh-5 overexpressors and the wild type tobacco seedlings were germinated and grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings at both 50 and 100 mM NaCl solution. But at 0, 150, and 200 mM NaCl concentration, the difference in growth rate was not detected.

• Plant Biotechnology Reports
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• 제5권1호
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• pp.19-25
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• 2011

Tocopherols belong to the plant-derived poly phenolic compounds known for antioxidant functions in plants and animals. Activation of mitogen-activated protein kinases (MAPK) is a common reaction of plant cells in defense-related signal transduction pathways. We report a novel non-antioxidant function of ${\alpha}$-tocopherol in higher plants linking the physiological role of tocopherol with stress signalling pathways. Pre-incubation of a low concentration of $50{\mu}M$ ${\alpha}$-tocopherol negatively interferes with MAPK activation in elicitor-treated tobacco BY2 suspension culture cells and wounded tobacco leaves, whereas pre-incubated BY2 cells with ${\alpha}$-tocopherol phosphate did not show the inhibitory effect on stimuli-induced MAPK activation. The decreased MAPK activity was neither due to a direct inhibitory effect of ${\alpha}$-tocopherol nor due to the induction of an inhibitory or inactivating activity directly affecting MAPK activity. The data support that the target of ${\alpha}$-tocopherol negatively regulates an upstream component of the signaling pathways that leads to stress dependent MAPK activation.

• 한국연초학회지
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• 제1권1호
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• pp.1-8
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• 1979

담배 (Va 115) 細胞의 液 培養에서 담배細胞의 多量生産을 할 수 있는 Tank 培養에 關한 基礎調査로 培地造成의 果를 調査하였으며 細胞增殖率에 큰 影響을 미쳤던 2,4-D와 無機燐酸의 果에 關하여 調査하였다. 1. 培地造成에서 細胞增殖率에 많은 影響을 미쳤던 要素는 무糖, 無機燐酸의 濃度,窒素源의 形態 및 植物홀몬,特히 2, 4-D의 有無였다. 2. 무糖의 最適濃度는 3%였으며 3%以上의 濃度에서는 多少 細胞增殖率이 좋은 듯 하였으나 그 差異는 크지 않아 3% 程度로 足하였다. 無機燐酸濃度는 LS 培地內의 無機燐酸의 約 2.5 培인 0.30mg/ml 일 때 細胞增殖率이 가장 좋았다. 3. 液 培地의 窒素源은 암모니아態窒素와 硝酸態窒素가 1 : 2 일 때가 가장 좋았고 窒素源이 암모니아태 만으로 使用하였을 때 細胞增殖率이 가장 낮았으며 硝酸態만 使用되었을 때는 암모니아태만 쓰였을 때 보다는 좋았으나 암모니아태와 硝酸態의 比가 1 : 2 일 때 보다는 떨어졌다. 4. 液 培地에 2,4-D 添加와 無機燐酸濃度를 높이면 細胞增殖率이 增加되는 機作을 調査하기 爲하여 呼吸率과 14C - glucose 吸收利用을 調査하였다. 細胞의 吸收率은 2, 4-D를 添加하면 增加되며 14C-glucose의 吸收는 培地內에 2, 4-D가 包含되거나 (0.2 ppm) 燐酸濃度가 높아지면(對照의 2.5培) 더욱 많았고, 吸收된 14C-glucose는 糖 상태보다 다른 形態,特히 amino 酸이나 有機酸으로 많이 变하였다.

• Journal of Plant Biotechnology
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• 제35권1호
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• pp.87-94
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• 2008

Porcine epidemic diarrhea virus (PEDV)는 돼지의 급성장염을 유발하여 설사 등의 증상을 일으키는 바이러스이다. 본 연구에서는 PEDV 항원단백질을 생산하는 담배 배양세포주를 개발하고자 하였다. PEDV에서 항원성이 알려진 스파크 단백질의 일부분을 암호화하는 유전자를 PCR로 합성하여 4종류의 형질전환 벡터를 제작하였다. 담배 배양세포 BY-2를 재료로 하여 Agrobacterium tumefaciens을 매개로 형질전환하였다. 선발배지 (MS salt, $KH_2PO_4$ 370 mg/L, 2,4-D 0.18 mg/L, Thiamin HCl 1 mg/L, kanamycin 100 mg/L, 침랙무 400 mg/L)에서 캘러스를 3주 간격으로 3개월 동안 계대배양하여 카나마이신 저항성 캘러스를 선발하였다. 선발된 캘러스를 대상으로 PCR 분석한 결과 형질전환 효율은 75% 이상이었으며 벡터당 40 여개 이상의 형질전환 배양세포주를 얻었다. 형질전환 배양세포주를 대상으로 Southern blot 분석하여 PEDV 유전자가 고구마 식물체의 게놈으로 안정적으로 도입되었음을 확인하였다. Northern blot 분석 결과 PEDV 스파크 단백질 유전자가 높은 수준으로 발현함을 확인하였으며 dot blot으로 PEDV 스파크 단백질 고생산 배양세포주를 선발하였다. 형질전환 담배 배양세포로부터 생산된 PEDV 항원단백질을 BALB/c 마우스에 경구투여 하여 면역활성을 조사한 결과 형질전환 세포주인 35S::SP1-M, 35S::SP2-M, 35S::SP4-M 세포주에서 1:10의 희석배수까지 바이러스 억제효과가 관찰되었다. 제작된 형질전환 벡터는 고구마와 같은 경구용 사료작물에 활용할 수 있을 것이다.

• 식물조직배양학회지
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• 제27권5호
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• pp.375-377
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• 2000

It is now generally accepted that a phosphoinositide cycle is involved in the transduction of a variety of signals in plant cells. In animal cells, the hydrolysis of phosphatidyl-4,5-bisphosphate catalysed by phosphatidylinositol - specific phospholipase C yields to D-myo-inositol - 1,4,5-trisphosphate and diacylglycerol, which are well known second messengers. The binding of InsP$_3$to a receptor located on the endoplasmic reticulum triggers a calcium release from the endoplasmic reticulum. We have detected and partially characterised key components of phosphoinositide signaling. First, tobacco microsomal fraction and plasma membrane PI-PLC. Consecutively, using a radioligand binding assay we have identified a $Ca^{2+}$ -dependent high affinity InsP$_3$binding site in microsomal membrane fraction vesicle preparation and then we have measured inositol-1,4,5-trisphosphate induced calcium release from tobacco microsomal fraction. These findings suggest that phosphoinositide signaling system is present and operates in the tobacco suspension culture.e.

• Journal of Ginseng Research
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• 제20권2호
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• pp.168-172
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• 1996

Ginsenoside Rb,, at a concentration of 10 $\mu\textrm{g}$/ml and over, initiated the cycle of oscillation of ion flux in erythrocytes after the cells had been treated with a protonophore, carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone (FCCP) and then with a $Ca^{2+}$ ionophore, A23,3,. Its action was similar to the additional portion of $Ca^{2+}$-ionophore or $Ca^{2+}$ ion to the erythrocytes. Effects of $Rg_1$ and Rf were different from that of Rb,. They did not induce the oscillation. They, however, increased the extracellular $K^{+}$ concentration and pH without returning to the initial state in the erythrocytes processed with FCCP and $A_{23187}$. We established that ginsenosides from 20-(5)-panaxatriol family induced the membrane hyperpolarization in erythrocytes, which was attenuated by the pretreatment of $Rb_1$, a major component of 20-(5)-panaxadiol.