• Journal of Plant Biotechnology
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• 제31권4호
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• pp.279-283
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• 2004

백합 (Lilium longiflorum cv. Georgia) 화분의 생장과 agro-infiltration에 의한 일시발현에 대한 물리적, 화학적, 생물적 요인의 영향을 분석하였다 화분을 배지에 섞기 위한 물리적 과정이나 agro-infiltration을 위한 진공작업과정은 정상적 화분생장을 위하여 최소화되는 것이 바람직한 것으로 나타났다. 비교적 넓은 범위에서의 온도 (19 to 27$^{\circ}C$)나 pH(5.0 to 8.0)에서 화분의 생장이 유사하게 진행되었으며 화학적 요인으로서의 cefotaxime (300mg/L), acetosyringone (800 $\mu$M), syringealdehyde (800 $\mu$M) 등의 처리는 화분의 생장에 영향을 나타내지 않았다. 그러나 kanamycin의 경우 매우 심한 생장저해현상을 보였는데 25mg/L의 농도에서도 저해현상을 보이는 경우도 있었다. GUS유전자의 화분발현시 acetosyringone(200-400$\mu$M)의 처리에 의하여 그 효율이 약간 향상되는 것으로 나타났으나 syringealdehyde의 경우에는 효과가 없었다. 짧은 시간 내의 agro-infiltration과정과 이어서 18 hr의 화분 및 박테리아의 동시배양으로서도 acetosyringone의 첨가에 상관없이 화분에서의 GUS 일시 발현결과를 얻을 수 있었다.

• 한국방재학회 논문집
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• 제8권5호
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• pp.103-109
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• 2008

사면에서의 강우는 경사와 토양의 물리적 성질에 따라 침투와 유출이 분리되게 되는데, 사면 해석에서는 강우가 모두 침투된다는 가정 하에 분석이 이루어져 지하수위가 과대 산정되는 경향이 있다. 따라서 본 연구에서는 서울지점의 강우분석에 이어 해석적 침투모형 중 적용성이 뛰어난 NRCS 모형과 Horton 모형을 이용하여 토양형에 따른 침투량과 유출량을 분리시킨 후, 토양을 분석하였다. 사면의 경사는 국내 설계기준에 의해 많이 적용되는 경사에 대한 단면을 구성하였으며, 산정된 침투량을 바탕으로 경사보정을 한 후 수치해석을 통해 침투거동 및 지하수위양상에 대해 분석하였다. 분석 결과에서 토양형이 D형에서 A형으로 갈수록 지하수위 변화양상이 크게 나타났다.

• 농업생명과학연구
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• 제45권6호
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• pp.193-199
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• 2011

An efficient transient expression system has been developed and characterized for the production of foreign genes in seedlings. The seedlings can be easily produced from commercial seeds used for vegetable sprouts. In principal, a chemical abrasive was employed to generate wounds in seedlings prior to vacuum-infiltration with Agrobacterium tumefaciens bearing the target gene. This optimized chemical wounding-assisted agro-infiltration process resulted in up to 15-fold increase in $\beta$-glucuronidase (GUS) enzyme activity. This procedure has been used efficiently to express hepatitis B surface antigen (HBsAg) protein in a transient mode. Therefore, seedlings with proper wounds can be suggested as a convenient tool for the production of useful recombinant proteins.

• The Plant Pathology Journal
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• 제24권3호
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• pp.296-304
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• 2008

The transient and rapid expression system of a foreign protein in planta is a very useful technique in biotechnology application. We have investigated optimum condition of Agrobacterium-infiltration technique in which expression level of foreign proteins were maximized without detrimental effects on plants using GFP and Cucumber mosaic virus 2b protein, which is known as an enhancer of gene expression and a suppressor of post-transcriptional gene silencing(PTGS). The optimum expression level of both RNA and protein of GFP with minimum leaf impairment was obtained at $OD_{600}$=0.2 of Agrobactrium inocula. The steady-state levels of GFP RNA and protein generally peaked at 3 and 7 days post-infiltration(dpi), respectively. In the presence of 2b, both the magnitude and duration of GFP expression was highly increased and we could detect GFP level until 17 dpi. On the other hands, the 2b-mediated higher accumulation of foreign proteins resulted in the repression of normal leaf growth, possibly due to the limitation of supply of energy or materials required for growth maintenance. Using this Agrobacterium-infiltration system with 2b and GFP, we tested a hypothesis for the threshold model of PTGS initiation. Four GFP transgenic lines of N. benthamiana, which shows different expression level of GFP were tested to determine the threshold level for PTGS initiation. Agrobacterium-infiltration of GFP into those GFP-transgenic plants resulted in the co-silencing of the transgenic GFP. It was found that very low concentration of Agrobacterium with GFP and GFP+2b($OD_{600}$=0.002-0.02) which could not phenotypically induce an additive GFP expression, was enough to trigger PTGS pathway in all GFP transgenic plants. This strongly indicates that each GFP-transgenic plant should be expressing the transgenic GFP at its own pre-determined level and there was no buffer zone of additive GFP-expression to the threshold. In other words, the PTGS seems to be immediately activated as a self-defensive mechanism if an internal balance of gene expression is broken.

• Journal of Microbiology and Biotechnology
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• 제33권4호
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• pp.463-470
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• 2023

This study confirmed the change in functional composition and alcohol-induced acute liver injury in Aloe arborescens after fermentation. An acute liver injury was induced by administration of ethanol (3 g/kg/day) to C57BL/6J mice for 5 days. A fermented A. arborescens Miller leaf (FAAL) extract was orally administered 30 minutes before ethanol treatment. After fermentation, the emodin content was approximately 13 times higher than that of the raw material. FAAL extract significantly attenuated ethanol-induced aspartate aminotransferase, alanine aminotransferase, and triglyceride increases in serum and liver tissue. Histological analysis revealed that FAAL extract inhibits inflammatory cell infiltration and fat accumulation in liver tissues. The cytochrome P450 2E1, superoxide dismutase, and glutathione (GSH), which involved in alcohol-induced oxidative stress, were effectively regulated by FAAL extract in serum and liver tissues, except for GSH. FAAL also maintained the antioxidant defense system by upregulating heme oxygenase 1 and nuclear factor erythroid 2-related factor 2 protein expression. In addition, FAAL extract inhibited the decrease in alcohol dehydrogenase and aldehyde dehydrogenase activity, which promoted alcohol metabolism and prevented the activation of inflammatory response. Our results suggest that FAAL could be used as a potential therapeutic agent for ethanol-induced acute liver injury.

• 원예과학기술지
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• 제29권3호
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• pp.267-272
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• 2011

${\beta}$-carotene hydroxylase는 carotenoids의 생합성 경로에서 astaxanthin 및 zeaxanthin 합성에 관여하는 주요 효소이다. Astaxanthin과 zeaxanthin은 비타민 A 전구체보다 항암효과가 더 높다고 보고되고 있다. 따라서 ${\beta}$-carotene hydroxylase 유전자(ChyB)를 토마토 잎에서 분리하고 binary 벡터에 클로닝 한 후 pIG121-ChyB-tom으로 명명하였다. 토마토 ChyB 유전자를 Agrobacterium-mediated infiltration 방법을 이용하여 생육 8주 된 담배 잎에 일시적 형질전환을 하였다. 감염 0, 1, 2, 3일 후 RT-PCR 한 결과 담배에서는 거의 발현되지 않았던 ${\beta}$-carotene hydroxylase 전사체가 감염 1일 후부터 증가하여 2일 후 최대 전사량을 보이고 3일 후부터 감소하는 경향을 나타내었다. 이 결과를 바탕으로 감염 후 0, 2, 3일째 담배 잎을 채취하여 항산화 효능을 측정한 결과, 감염 2일 후 1,1-diphenyl-pricryl hydrazyl(DPPH) radical 소거 활성이 대조구에 비해 약 30% 증가하는 것을 확인하였다. 이 연구결과를 통해 외래 유용 유전자를 원예작물에 형질전환하여 항산화 및 항암효과가 우수한 zeaxanthin 및 astaxanthin 등 oxy-carotenoids의 함유량을 증진시킨 새로운 기능성 토마토 및 다양한 원예작물의 신품종 개발에 이용될 수 있을 것으로 기대된다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2003년도 추계학술대회
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• pp.195-195
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• 2003

The recruitment and infiltration of monocytes into the artery wall is a crucial early step in atherogenesis. KR-31378 has been shown to be a neuroprotective agent in rat brain via its potent antioxidant and antiapoptotic actions. Here, we report the effects of this compound on atherogenesis, and some possible mechanisms of action.(omitted)

• The Plant Pathology Journal
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• 제21권2호
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• pp.158-163
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• 2005

Virus-induced gene silencing (VIGS) is a recently developed gene transcript suppression technique for characterizing the function of plant genes. However, efficient VIGS has only been studied in a few plant species. In order to extend the application of VIGS, we examined whether a VIGS vector based on TRV would produce recognizable phenotypes in soybean. Here, we report that VIGS using the Tobacco rattle virus (TRV) viral vector can be used in several soybean cultivars employing various agro-inoculation methods including leaf infiltration, spray inoculation, and agrodrench. cDNA fragments of the soybean phytoene desaturase(PDS) was inserted into TRV RNA-2 vector. By agrodrench, we successfully silenced the expression of PDS encoding gene in soybean. The silenced phenotype of PDS was invariably obvious 3 weeks after inoculation with the TRV-based vector. Real-time RT-PCR analyses showed that the endogenous level of GmPDS transcripts was dramatically reduced in the silenced leaf tissues. These observations confirm that the silenced phenotype is closely correlated with the pattern of tissue expression. The TRV-based VIGS using agrodrench can be applied to functional genomics in a soybean plants to study genes involved in a wide range of biological processes. To our knowledge, this is the first high frequency VIGS method in soybean plants.

• 생태와환경
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• 제52권4호
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• pp.394-402
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• 2019

The Agrobacterium tumefaciens mediated gene transfer is widely used to generate genetic transformation of plants and transient assay of temporal exogenous gene expression. Syringe infiltration system into tobacco (Nicotiana benthamiana) leaves is a powerful tool for transient expression of target protein to study protein localization, protein-protein binding and protein production. However, the protocol and technical information of transient gene expression, especially double strand RNA (dsRNA), in tobacco using Agrobacterium is not well known. Recently, dsRNA is crucial for insecticidal effect on destructive agronomic pest such as Corn rootworm. In this study, we investigated the factor influencing the dsRNA expression efficiency of syringe agro-infiltration in tobacco. To search the best combination for dsRNA transient expression in tobacco, applied two Agrobacterium cell lines and three plant vector systems. The efficiency of dsRNA expression has estimated by real-time PCR and digital PCR. As a result, pHellsgate12 vector constructs showed the most effective accumulation of dsRNA in the cell. These results indicated that the efficiency of dsRNA expression was depending on the kind of vector rather than Agrobacterium cells. In summary, the optimized combination of transient dsRNA expression system in tobacco might be useful to in vivo dsRNA expression for functional study and risk assessment of dsRNA.

• 생명과학회지
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• 제31권8호
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• pp.736-744
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• 2021

강화사자발쑥(Artemisia princeps Pampanini)은 항산화, 항염증 및 항균제와 같은 면역 기능 관련 질병에 널리 사용되는 약초이다. 이 연구에서 우리는 RAW 264.7 세포에서 AP 추출물의 항염증 효과를 조사하고 관련 메커니즘을 평가하였다. AP 추출물의 효과는 complete Freund's adjuvant (CFA) 유도 관절염 및 lipopolysaccharide(LPS) 유도 마우스 모델에서도 평가되었다. RAW 264.7 세포에서 AP 추출물은 LPS에 의해 유도 된 산화질소(NO) 생성과 inducible NO synthase 및 cyclooxygenase-2 단백질 발현을 현저하게 억제했다. RAW 264.7 세포에서 LPS로 유도된 mitogen-activated protein kinase 와 nuclear factor-κB의 인산화 또한 AP 추출물에 의해 유의하게 억제되었다. AP 추출물의 경구 투여는 CFA 처리 마우스 그룹에 비해 발의 부종 및 비장 지수 증가를 억제하였다. 조직학적으로 CFA 처리 마우스 군에서는 cartilage와 synovium에서 염증 세포의 침윤이 증가한 반면 AP 추출물 투여군에서는 억제되었다. 더욱이, AP 추출물은 염증성 사이토카인으로 알려진 tumor necrosis factor-α 수준을 CFA 및 LPS 처리 마우스 모델에서 현저하게 감소시켰다. 결론적으로, AP 추출물의 항염증 및 항 관절염 효과는 in vitro 및 in vivo 모델 모두에서 확인되었으며, 이는 Artemisia princeps Pampanini가 관절염 치료의 후보 물질이 될 수 있음을 시사한다.