• 식물조직배양학회지
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• 제25권1호
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• pp.45-50
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• 1998

꽃양배추의 하배축 조직을 proteinase inhibitor II 유전자가 도입된 Agrobacterium tumefaciens LBA 4404와 2일간 pH 5.5로 조절된 MS 액체배지에서 공동배양후 carbenicillin 500mg/L kanamycin 20mg/L와 BA 1mg/L가 함유된 MS 재분화배지에 옮겼다. 이들 조직을 매 2주마다 계대배양하였으며 약 4주후에 kanamycin 저항성 개체를 얻었다. 형질전환된 것으로 추정되는 식물체는 kanamycin 30mg/L가 함유된 선발배지에서 생존하였다. PCR 분석결과, PI-II 유전자가 형질전환체의 게놈상에 삽입되어 있음을 확인하였다. 형질전환체의 Southern blot 분석을 통하여 ECL-labelling된 PI-II 유전자와 동일한 것으로 판단되는 약 500bp 위치에서 밴드를 확인할 수 있었다.

• 식물조직배양학회지
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• 제25권2호
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• pp.95-98
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• 1998

PI-II cDNA가 도입된 식물발현 벡터인 pGA875를 가진 A.tumefaciens LBA4404를 이용하여 꽃양배추의 하배축 조직에 형질전환하여 식물체를 재분화 시켰다. Dot blot 분석으로 PI-II 유전자가 전사됨을 확인할 수 있었다. 또한 이들 개체를 담배거세미나방 유충을 이용하여 생물검정한 결과 대조구에 비해 형질전환체 잎의 섭식정도가 현저히 떨어지는 것을 알 수 있었다. 개화후 이들 개체의 종자를 받아 후대검정을 실시하였을 때 27,4%가 kanamycin내성을 가진 꽃양배추로 확인되었다.

• Applied Biological Chemistry
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• 제38권5호
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• pp.382-386
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• 1995

감자의 단백질 분해효소 억제제-II(PI-II)단백질은 카이모트립신 저해부위와 트립신 저해부위로 나뉘어 있는데 PI-II 유전자중의 하나인 PI-IIT 유전자의 염기서열에서 비롯된 아미노산 서열이 폴리펩타이드 카이모트립신 저해제(PCI) 단백질의 아미노산 서열과 84%의 높은 동질성을 가지고 있으므로 감자의 단백질 분해효소 억제제유전자 집단 (family) 중의 하나인 PCI와 homology가 있는 유전자단편을 클로닝하기 위하여 이미 클론된 PI-IIT 유전자로부터 PCR을 행하여 얻어진 DNA 단편을 백터에 클로닝하고 염기서열을 결정하였다. 염기서열을 확인한 유전자 단편을 박테리오파아지 T7 promoter와 terminator를 갖고있는 플라스미드 pET3a에 옮겨 대장균 BL2l(DE3)에서 발현시켰던바 IPTG의 부가에 따라 유기되는 것을 확인 하였으나 발현수준은 기대했던것에 미치지 못하였다.

• 식물조직배양학회지
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• 제25권1호
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• pp.37-43
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• 1998

꽃양배추 '은배' 종자를 무균 파종한 후 6일째된 하배축 조직을 BA 1㎎/L, sucrose 30㎎/L한천 8 g/L가 첨가된 MS 재분화배지에 1일간 전처리한 다음, PI promoter-GUS 융합 유전자가 도입된 Agrobacterium tumefaciens LBA 4404와 2일간 동일조성의 MS 액체배지에서 공동배양하여 carbenicillin 500 ㎎/L와 kanamycin 20 ㎎/L가 첨가된 MS 재분화배지로 옮겨 주었을 때 가장 많은 형질전환체를 얻을 수 있었다. PCR 분석결과, PI promoter-GUS 융합 유전자가 형질전환체의 게놈상에 삽입되어 있음을 확인하였다. Southern 분석결과, ECL-labelling된 PI promoter-GUS 융합 유전자 probe의 coding sequence와 동일한 것으로 판단되는 약 366bp 위치에서 밴드를 확인할 수 있었다. 그러나 형질 전환되지 않은 식물체에서는 이들 밴드를 확인할 수 없었다. 조직내 GUS 유전자의 활성은 잎부위에서부터 시작하여 엽병과 줄기의 관다발을 중심으로 나타났으며 상처의 정도가 심할수록 높은 편이었고 그 범위도 넓었다.

• Plant Resources
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• 제6권3호
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• pp.205-210
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• 2003

Ginseng(Panax ginseng C.A. Meyer) is a perennial herbaceous plant which grows very slowly. It takes about 3 to 4 years from seeding to collecting the ripe seeds and the ginseng propagation is very difficult. and so, it is very difficult to breed ginseng plant. Ginseng tissue culture was started from at 1960, and ginseng commercial product by in vitro callus culture was saled, however upto now, regenerants were not planted to soil normally. Recently, plant genetic engineering to produce transgenic plants by introducing useful genes has been advanced greatly. In a present paper, transformation of ginseng plants was achieved by co-cultivation with Agrobacterium harboring the binary vector coding Proteinase-II gene, which confer resistant or tolerant to insect pests, The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 35S promoter. The NPT II gene and introduced genes of the transgenic ginseng plants were successfully identified by the PCR. Especially the transgenic ginseng plants were regenerated using new techniques such as repetitive single somatic embryogenesis.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Signal transduction in Toxicology
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• pp.96-106
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• 2001

The phase II detoxifying enzymes are inducible by a variety of compounds and play an essential role for the protection of cells. Many of chemoprotective agents trigger cellular signals for the phase II enzyme induction, which subsequently activate gene transcription through ARE activation.(omitted)

• 자연과학논문집
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• 제10권1호
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• pp.33-37
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• 1998

식물의 주요 phytohormone의 하나인 cytokinin은 식물체의 줄기와 뿌리성장 그 외에도 영양의 전달이나 노화방지, 열매숙성 등 식물의 성장과 발달에 미치는 영향은 크고 다양하다. Cytokinin 생합성에 관여하는 효소를 생산하는 것으로 알려져 있으며 토양박테리아 Agrobacterium tumefaciens에 존재하는 유전자인 isopentenyl transferase (jpt)를 이용한 많은 분자생물학적 연구가 진행되어 왔는데 그 중 하나로 이 연구에서 jpt 유전자에 의한 cytokinin의 overproduction이 식물체에 성장과 발달에 어떠한 영향을 주는지 관찰하고 그 결과가 제시할 수 있는 작물의 유전 공학적 이용가능성에 대하여 알아본다.

• Toxicological Research
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• 제17권
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• pp.251-256
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• 2001

The expression of phase II detoxifying enzymes is affected by a variety of compounds and the induction of the enzymes plays an essential role in chemoprevention. A variety of phytochemicals such as sulfur-containing chemoprotective agents (SCC) may trigger cellular signals and activate phase II gene expression through ARE activation. see induces glutathione S-transferases. Studies were conducted to investigate the role of mitogen-activated protein (MAP) kinase and phosphatidylinositol 3-kinase (PI3-kinase) in the induction of GST (e.g. rGSTA2) by sec. We also studied the MAP kinase pathway responsible for the GST expression by see and compared that with the pathway activated by oxidative stress as a result of sulfur amino acids deprivation (SAAD). see inhibited phosphorylation of ERK1/2 although the effect of see on JNK and p38 MAP kinase was minimal. Wortmannin and LY294002. PI3-kinase inhibitors. abolished the increases in rGSTA2 mRNA and protein levels by SCC. Deprivation of cystine and methionine caused oxidative stress in H4IIE cells. as evidenced by a decrease in the reduced glutathione and an increase in prooxidant production. Electrophoretic mobility shift assay revealed that the ARE complex consisting of Nrf-1/2 and Maf proteins was activated 12~48 h. The rGSTA2 mRNA and protein levels were increased by SAAD. Activation of ARE and induction of rGSTA2 were both completely inhibited by PI3-kinase inhibitors. Inhibition of p38 MAP kinase by SB203580 prevented the ARE-mediated rGSTA2 induction. The results of this study showed that PI3-kinase might play an essential role in the ARE-mediated rGSTA2 induction by see or SAAD and that the dual MAP kinase pathways were responsible for the enzyme induction.

• 대한예방한의학회지
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• 제22권1호
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• pp.107-127
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• 2018

Objectives : The present study investigated the anti-obese and blood flow improvement activities of aqueous extracts of ginseng berry (GBe) on the mild diabetic obese mice as compared with metformin. Methods : After end of 56 days of continuous oral administrations of GBe 150, 100 and 50 mg/kg, or metformin 250 mg/kg, anti-obese and blood flow improvement effects - the changes of body weights, body and abdominal fat density by in live dual-energy x-ray absorptionmetry (DEXA), tail bleeding time, prothrombin time (PT), activated partial thromboplastin time (aPTT), serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and high density lipoprotein (HDL) levels, aorta and serum cyclic guanosine monophosphate (cGMP), nitric oxide (NO) and endothelin (ET)-1 levels, aorta phosphorylated PI3K (pPI3K), phosphorylated Akt (pAkt) and phosphorylated p38 MAPK (pp38 MAPK) levels were systemically analyzed. In addition, aorta vascular dilation and constriction related gene mRNA expressions - PI3K, Akt, eNOS, p38 MAPK and ET-1 were also analyzed by realtime RT-PCR. Results : The obesity and related blood flow impairment, induced by 84 days of continuous HFD supply, were significantly inhibited by 56 days of continuous oral treatment of GBe 150, 100 and 50mg/kg, dose-dependently, and they also dramatically normalized the changes of the aorta vascular dilation and constriction related gene mRNA expressions, also dose-dependently. Especially, GBe 150 mg/kg constantly showed favorable inhibitory activities against type II diabetes related obesity and vascular disorders through PI3K/Akt pathway and p38 MAPK mediated cGMP, NO and ET-1 expression modulatory activities, as comparable to those of metformin 250 mg/kg in HFD mice. Conclusion : By assessing the key parameters for anti-obese and blood flow improvement activities on the HFD-induced mild diabetic obese mice, the present work demonstrated that GBe 150, 100 and 50 mg/kg showed favorable anti-obese and blood flow improvement effects in HFD-induced type II diabetic mice, through PI3K/Akt pathway and p38 MAPK mediated cGMP, NO and ET-1 expression modulatory activities.

• Toxicological Research
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• 제23권2호
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• pp.127-133
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• 2007

Metformin is a drug used to lower blood sugar levels in patients with type 2 diabetes via activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK). The primary objective of this study was to investigate whether metformin at the pharmacologically effective concentrations affects the expressions of ${\gamma}$-glutamylcysteine synthetase and phase II antioxidant genes in the H4IIE cell. Treatment of the cells with either metformin or 5-aminoimidazole-4-carboxamide riboside (AICAR) abrogated tert-butylhydroxyquinone (t-BHQ) induction of ${\gamma}$-glutamylcysteine synthetase, a rate limiting enzyme of GSH synthesis. The ability of t-BHQ to induce glutathione S-transferases (GSTs), a major class of phase II detoxifying enzymes that playa critical role in protecting cells from oxidative stress or electrophiles, was also inhibited by the agents. Transcriptional gene repression by metformin was verified by the GSTA2 promoter luciferase assay. Moreover, either metformin or AICAR treatment significantly decreased t-BHQ-dependent induction of other GSTs (i.e., $GST{\mu}$ and $GST{\pi}$ forms). Taken together, our data indicate that metformin treatment may result in the repression of ${\gamma}$-glutamylcysteine synthetase and glutathione S-transferase genes possibly via AMPK activation.