• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.239-243
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• 2000

Helicobacter pylori, an etiologic agent of gastritis and peptic ulceration, produces urease which elicits a powerful immunoglobulin response in H. pylori-infected individuals. To establish a model plant vaccine agains H. pylori, 750 bp -ureA DNA amplified by polymerase chain reaction from pH 808 plasmid harboring urease gene cluster was cloned and manipulated to be expressed in tobacco plants. From the regenerated transgenic tobacco plants, ureA DNA integration,m its mRNA expression and protein synthesis were analyzed and confirmed by standard molecular techniques. The CaMV 35S promoter-driving ureA construct was expressed to produce a 30 kDa protein which was identical with bacterial UreA in size when detected on immunoblot of SDS polyacrylamide gel electrophoresis.

• Research in Plant Disease
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• v.21 no.3
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• pp.186-192
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• 2015

Cucumber mosaic virus possesses 2b gene known as a suppressor of post-transcriptional gene silencing (PTGS). To investigate its function and effect in plant, transgenic Nicotiana benethamiana expressing 2b gene was developed and analyzed in phenotypic characteristics and differential gene expression (DEG) comparing with wild-type. Eight lines of transgenic plants ($T_0$) were obtained with difficulty and showed severe deformed phenotypes in leaves, flowers, petioles and etc. Moreover, transgenic plants were hardly able to set seeds, but small amounts of seeds were barely produced in some of transgene-hemizygous plants. DEG analysis showed that transgenic plant ectopically accumulated diverse RNA transcripts at higher levels than wild-type probably due to the disturbance in RNA metabolism, especially of RNA decay, caused by 2b-mediated inhibition of PTGS. These ectopic accumulations of RNAs disrupt protein and RNA homeostasis and then subsequently lead to abnormal phenotypes of transgenic plants.

• Journal of Life Science
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• v.18 no.8
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• pp.1023-1030
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• 2008

NtMEK2, which is the tobacco MAPK kinase that is upstream of SIPK and WIPK, was identified using the dexamethasone (DEX)-inducible gain-of-function transgenic system. Expression of $NtNEK2^{DD}$, a constitutively active mutant of NtNEK2, leads to HR-like cell death, which indicates that the NtMEK2-SIPK/WIPK cascade controls defense responses in tobacco. However, little is known about the downstream target substrates or defense-related genes that are regulated by the NtMEK2-SIPK/ WIPK cascade. In this study, ACP-based differential display RT-PCR was used to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade in $NtNEK2^{DD}$ transgenic plants. The results identified 6 novel differentially expressed genes (DEGs). These included pathogen induced protein 2-4 (pI2-4), monoterpene synthase 2 (MTS2), seven in absentia protein (SINA), cell death marker protein 1 (CDM1), hydroxyproline-rich glycoprotein (HRGP) and unknown genes (DEG45). The induction of these genes was confirmed by RT-PCR of samples obtained from $NtNEK2^{DD}$ plants. Additionally, when compared with other isolated DEGs, the pI2-4, CDM1 and HRGP genes were significantly up-regulated in response to treatment with salicylic acid and tobacco mosaic virus. Taken together, these results suggest that three novel DEGs were regulated by the NtMEK2-SIPK/WIPK cascade involved in disease resistance in tobacco.

• Journal of Plant Biotechnology
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• v.32 no.2
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• pp.97-103
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• 2005

Antioxidative responses of transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts was investigated with several herbicides. In greenhouse test, tolerance of SOD/APX-overexpressed tobacco (CA) to photosystem (PS) I inhibitor paraquat was increased by about 40%. However, any response differences between CA and wild type (WT) tobacco was not observed in a treatment with PS II inhibitors (bromoxynil, diuron and bromacil), chlorophyll biosynthesis inhibitor(oxyfluorfen), carotenoid biosynthesis inhibitor (fluridone) and 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase inhibitor (glyphosate). This tendency was also similar in the growth chamber test of low light intensity, using paraquat and diuron. That is, increased antioxidant activity of CA was shown only in paraquat treatment. When paraquat was foliar-treated to 6 to 9-leaf stage plant, the third to fourth placed leaf from shoot tip showed relatively higher antioxidant activity. Ascorbate supplemented to paraquat solution alleviated the phytotoxicity with a similar range in both CA and WT. In conclusion, CA specifically responded to oxidative stress induced by paraquat among tested herbicides in a whole plant assay.

• Applied Biological Chemistry
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• v.37 no.5
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• pp.343-348
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• 1994

Ti plasmid of A. tumefaciens was labeled with $^3H-thymidine$, purified and encapsulated into phosphatidylserine (PS) and PS-cholesterol (Chol; 1 : 1 molar ratio) liposomes by lyophilization-rehydration method. PS was supplemented with 1 mole percent octadecyl rhodamine B for fluorometric measurement of PS. Liposomes entrapping $^3H-Ti plasmid$ were fused with Nicotiana sanderae protoplasts by treating with 5 mM $CaCl_2$ and 10% PEG. The fusion was evidenced by fluorescence microscopic technique. The amounts of Ti plasmid and PS associated with protoplasts were assayed by the radioactivity of $^3H-Ti plasmid$ and by the fluorescence of rhodamine B. About 7.9% of the PS liposome and 7.2% of PS-Chol liposome were fused with protoplasts. During the fusion process, about 30% of the liposomal contents of PS-Chol liposome was leaked, in contrast to about 60% leakage of its contents in PS liposome. Accounting the number of liposomes fused with protoplasts together with the encapsulation efficiency and the leakage of liposomal contents, it was calculated that ca. 1,700 Ti plasmid was transfered into one protoplast by the present method. This result may indicates that the present method transfers enough Ti plasmid into plant protoplast to elicit genetic transformation of plants.

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2002.05a
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• pp.3-42
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• 2002

21세기에는 인구의 폭발적 증가와 함께 가속화된 산업화로 말미암아 경지 면적은 줄고 농업 환경은 더욱 피폐해질 것으로 예상된다. 지금도 이미 화석 에너지원의 고갈로 대체 에너지 개발이 시급히 요구되고 있으며, 지구의 자연 환경 보존 목소리도 그 어느 때 보다 높다. 한마디로 식량, 에너지, 환경 문제가 새 세기에 우리가 시급히 해결해야할 과제로 주어져 있다. 이에 과학계에서는 식량 및 대체 에너지원의 공급을 증대시키고 환경을 보존할 수 있는 보편적인 수단으로 환경 친화적 유전자 변형 (GM)작물의 활용이 제시되고있다. 따라서 선진국들은 이의 기반이 되는 식물유전체 연구에 대규모 투자를 아끼지 않고 있으며, 이를 이 용한 식물 생명공학산업을 국가 전략 산업으로 집중 육성하고 있다. GM작물 제조 기술은 유용 유전자의 발굴 및 재조합, 식물세포로의 이식 및 재분화를 통한 완전한 식물체 재생, 이를 품종으로 실용화하는 단계로 구성되어 있다. GM작물은 1983년 항생제 저항성 담배가 개발된 것을 시점으로 하여, 1994년에는 연화지 연 토마토 Flaver Saver이후 지금까지 개발 실용화된 작물은 제초제 저항성 콩, 카놀라, 목화, 그리고 해충 저항성 옥수수 등이 있으며,2001년까지 세계적으로 상품화 승인을 얻은 경우는 15 작물 68품종에 이른다. 2001년 경우 GM작물 종자시장은 약 30억 달러에 달하고 있으며, 미국, 아르헨티나, 캐나다 등 세계적으로 52.6백만 ha에 이르는 면적에서 재배되었다. 그러나 GM농산물의 식품 및 환경 안전성에 대한 의구심이 일기 시작하였고, 따라서 이의 생산 및 소비에 대한 전반적 인 문제가 뜨거운 쟁점으로 부각되기도 하였다. 이 에 각국 정부는 객관적 인 안전성을 확보하기 위한 제도적인 장치를 마련하고 있으며, 아울러 과학기술자들은 더욱 안전한 형질전환 기술 개발을 도모하고 있다. 다음 세대의 GM작물은 단순한 제초제 및 병해충 저항성을 넘어서서 특정 영양 또는 건강기능성을 향상시켜 부가가치를 증가시킨 신품종 맞춤작물이 지속적으로 개발 상업화될 것이다. 따라서 고유성을 가진 유용 유전자의 대량 확보 여부가 산업적 경쟁력을 결정하게 될 것이다. 지금까지 개별 유전자 중심으로 이루어지던 유용 유전자 발굴 작업은 유전체학의 출현으로 규모가 대량화되고 그 효율이 증진되었다. 따라서 진 각국은 유용 유전자 발굴에 국가적 차원의 역량을 집중하고 있다. 그러나 우리나라는 정부와 민간의 소규모 지원으로 근근히 기술 습득 차원에 머물러 왔으며, 산업적 경쟁력의 무기가 될 고유한 유용 유전자와 형질전환 기술이 거의 없는 어려운 상황에 놓여 있다. 최근 정부가 시작한 생명공학 분야 대규모 연구지원 사업 기대를 모아 보며 이 분야 과학기술자들의 노력을 촉구한다.

• Korean Journal of Soil Science and Fertilizer
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• v.46 no.1
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• pp.58-64
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• 2013

The objective of this study was to understand the physiological response and cadmium accumulation of MuS1 transgenic tobacco exposed to high concentration of Cd in soil. For this, a pot experiment was carried out in a greenhouse for a month, with two lines of MuS1 transgenic tobaccos (S4 and S6) and non-transgenic tobacco cultivated in the soils spiked at three different Cd concentrations (0, 60 and 180 mg $kg^{-1}$). Both transgenic and non-transgenic tobacco showed visible toxic symptoms such as chlorosis and leaf roll as treated concentration increased. The net photosynthetic rates of MuS1 plants (S4 and S6) exposed at 180 mg $kg^{-1}$ Cd were 6.3 and $7.7{\mu}mol\;m^{-2}s^{-1}$, being higher than those of the non-transgenic plant ($4.8{\mu}mol\;m^{-2}s^{-1}$). Values of stomatal conductance of MuS1 transgenic plants (0.05 and 0.008 mmol $H_2O\;m^{-2}s^{-1}$) were also higher than those of non-transgenic plant (0.03 mmol $H_2O\;m^{-2}s^{-1}$). In addition, fresh and dry weights of MuS1 transgenic plants were heavier than those of non-transgenic plant. Likewise, MuS1 transgenic plants appeared to be better physiological performance than non-transgenic tobacco when exposed at high concentration of Cd in soil. With regard to metal accumulation, MuS1 transgenic tobaccos accumulated more Cd in their roots than non-transgenic tobacco implying that MuS1 transgenic tobacco is suggested to be used for phytostabilization of heavy metals.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.41-45
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• 2008

Laccase (EC 1.10.3.2) is a small group of enzymes that catalyze the oxidation of a broad range of phenolic compounds including hazardous and recalcitrant pollutants in the environment. This study attempted to develop an efficient system for production of a recombinant laccase by chloroplast genetic transformation of tobacco. Chloroplast transformation vector was constructed and introduced into the tobacco chloroplast genome using particle bombardment. Chloroplast-transformed plants were subsequently regenerated. PCR and southern blot analyses confirmed stable integration of the laccase gene into the chloroplast genome. Northern blot analysis revealed that mRNA of the laccase gene was highly expressed in chloroplast-transformed plants.

• The Journal of Natural Sciences
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• v.10 no.1
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• pp.33-37
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• 1998

Cytokinin is one of major growth regulators in plants. In this study, the gene isopentenyl transferase (jpt) which encodes a key enzyme involved in the biosynthesis of the growth regulator cytokinin isolated from Agrobacterium tumefaciens was introduced ito tobacco plant via Agrobacterium-mediated transformation. The jpt gene was modulated using the proteinase inhibitor II (PI-IIK) promotor. In general, this promoterlipt gene fusion resulted in overproduction of cytokinins throughout the transgenic plants. The overproduction of cytokinin caused dramatic changes in morphology of the plant, including stem thickness and reduced root development. The studies reported in this paper were initiated to examine the consequences of overproduction of cytokinin in plant.

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.201-205
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• 1995

The synthetic oligomers called nos right palindrome (RP) element and left palindrome (LP) element were inserted into nos.minimal promoter nos 5'-101 deletion mutant The activity of nos promoter was measured by studying the expression pattern of gene fusion between nos promoter and reporter genes such as chloramphenicol acetyltransferase and $\beta$-glucuconidase. Analysis of transgenic tobacco plane carrying transgene showed that the activity of nos minimal promoter activity was recovered by insertion of synthetic nos RP element. Nos RP element insertion of nos minimal promoter was induced by auxin, dithiothreitol, salicylic acid and methyl jasmonate.