• Title/Summary/Keyword: Selective vector

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Transgenic Sweetpotato (Ipomoea batatas) Expressing Spike Gene of Porcine Epidemic Diarrhea Virus (돼지 유행성 설사병 바이러스의 스파이크 유전자 발현 형질전환 고구마)

  • Yang Kyoung-Sil;Lim Soon;Kwon Suk-Yoon;Kwak Sang-Soo;Kim Hyun-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.263-268
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    • 2005
  • Porcine epidemic diarrhea virus (PEDV) causes acute enteritis in pigs of all ages and is often fatal for neonates. In order to develop sweetpotato plants expressing PEDV antigen, we constructed the vector expressing spike gene of PEDV under the control of sweetpotato sporamin promoter or constitutive CaMV 35S promoter. The spike protein region of PEDV was synthesized by PCR and linked to each promoter, Transgenic sweetpotato [Ipomoea batatas (L.) Lam. cv. Yulmi] plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. The co-cultured embryogenic calli transferred to selective MS medium containing 1 mg/L 2,4-D, 100 mg/L kanamycin, and 400 mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 3 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the spike gene of PEDV was inserted into the genome of the sweetpotato plants. RT-PCR revealed that the spike gene of PEDV was highly expressed in transgenic sweetpotato plants.

Expression of CP4 5-Enol-Pyruvylshikimate-3- Phosphate Synthase Transgene in Inbred Line of Korean Domestic Maize (Zea may L.) (국내 옥수수 순계주에서 CP4 5-Enol- Pyruvylshikimate-3- Phosphate Synthase 유전자의 발현)

  • Cho, Mi-Ae;Kwon, Suk-Yoon;Kim, Jin-Seog;Lee, Byoung-Kyu;Moon, Choo-Yeun;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.375-380
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    • 2007
  • This study was conducted to develop herbicide-resistance domestic maize plants by introducing the CP4 5-enol-pyruvylshikimate-3-phosphate synthase (CP4 EPSPS) gene using Agrobacterium tumefaciens-mediated immature embryo transformation. Immature embryos of five genotypes (HW1, KL103, HW3, HW4, HW7) were co-cultivated with strains Agrobacterium tumefaciens (strain C58C1) containing the binary vector (pCAMBIA2300) carrying Ubiquitin promoter-CP4 EPSPS gene and Cauliflower mosaic virus 35S (CaMV35S) promoter-nptll gene conferring resistance to paromomycin as a selective agent. The presence and expression of CP4 EPSPS transgene were confirmed by PCR, RT-PCR and Northern blot analysis, respectively. Also, the resistance to glyphosate in the transgenic maize ($T_1$) was analyzed by shikimate accumulation assay. The frequency (%) of paromomycin-resistance callus was 0.37, 0.03, 2.20, 2.37, and 0.81% in pure lines HW1, KL103, HW3, HW4 and HW7, respectively. EPSP transgene sequences were amplified in putative transgenic plants that regenerated from paromomycin-resistance calli of two inbred lines (HW3, HW4). Of them, RT-PCR and Northern blot analyses revealed that the transgene was only expressed in two transgenic events (M266, M104) of HW4 inbred line, and a mild glyphosate resistance of transgenic event (M266) was confirmed by the lower shikimate accumulation in leaf segments. These results demonstrate that transgenic maize with herbicide-resistance traits in Korean genotype can be genetically obtained.

Experimental study for Selective Withdrawal on Stratified Water Tank by using PIV (PIV를 활용한 성층수조에서 선택취수방안에 대한 실험적 연구)

  • Son, Byung-Ju;Park, Jae-Hyeon;Kim, Young-Do
    • Proceedings of the Korea Water Resources Association Conference
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    • 2007.05a
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    • pp.556-559
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    • 2007
  • 고탁수 장기화 문제를 해결하기 위한 방법으로는 고탁수층을 선택취수하여 우선 배제함으로써 하류하천의 고탁수 발생일수를 최소화하는 방법이 있는데, 이와 같은 선택적 취수기법은 저수지 운영에 있어서 고탁수층을 우선 배제한 후 홍수기 이후에 저수지내로 유입되는 청수를 담수하여 호소 내 탁도 저감 효과를 얻을 수 있다. 본 연구에서는 선택취수 시 성층수괴의 동수역학적 변화를 분석하기 위하여 국내에서 처음으로 Two-tank 기법을 이용하여 성층구조를 구현해 내었는데, 소금물의 밀도변화를 이용하여 수심 1m의 성층수조를 만들었고, 밀도경사가 상이한 성층구조에서 취수조건을 변화시키면서 비교란 유속계인 PIV 시스템을 이용해 유속의 흐름을 Vector와 Contour로 분석하였다. 선택취수에 대한 흐름의 동수역학적 분석결과 취수유량보다는 밀도성층경사 변화에 더 민감한 반응을 보였다. 취수유량을 줄이거나 밀도성층경사를 급하게 했을 때에는 선택취수 영역(withdrawal zone)의 수직방향 폭은 줄어드는 반면, X축으로의 영향범위는 증가함을 나타냈다. 취수유량을 증가시키거나 성층밀도경사를 완만히 했을 땐 선택취수 영역(withdrawal zone)의 수직방향 폭은 증가하였고 X축으로의 영향범위는 축소됨을 나타내었다. 이 결과는 Richardson 수로도 판단되어지는데, Richardson 수가 증가하면 유속에 비해 상대적으로 밀도성층경사가 크다는 것인데 이럴 경우 선택취수 영역(withdrawal zone)의 수직방향 폭 최대가 되고 선택취수 영역(withdrawal zone)의 수직방향 폭은 최소가 된다. 선택취수 영역(withdrawal zone)의 수직방향 폭이 최소가 되면 취수구 직경 D의 1.8배의 값을 가지고 Richradson 수가 최소가 되더라도 취수구 직경 D의 3.3배를 벗어나지 않는다는 결과를 도출할 수 있었다.링 목적으로 사용될 수 있다. 본 연구에서 개발한 영상수위계는 한강홍수통제소 관할의 전류, 청담대교 등 4개소 낙동강 홍수통제소 2개소, 지자체 등에 적용되었으며, 적용 결과 비교적 안정적이면서 정확하게 수위를 측정하는 것으로 나타났다. 한편 기존 CCD 카메라 이외에 CCTV를 이용한 영상수위계를 개발하여 영상의 화질 개선뿐 아니라 하천화상 감시 기능을 강화하였다.소류의 섭취율은 높았다. 집단간의 상관도를 보면 교육별로 김치, 장아찌, 콩이 각각 p>0.5 수준에서 유의한 차가 없었고, 나머지는 유의한 차가 있었다. 연령별로는 멸치가 유의한 차가 없었고(p>0.5), 수입별로는 콩이 유의한 차가 없었다(p>0.5). 4. 영양지식(營養知識) 검토 가정생활(家庭生活)에 필요(必要)한 일반적(一般的)인 영양지식(營養知識)은 대체적으로 낮은 편이었다. 어린이 영양, 편식의 해로움, 비만증의 해로움, 임신부 그리고 수유부 영양에 대하여는 일반적으로 알고 있다고 하였으며, 그다음으로 이유기 영양, 어린이 발육에 필요한 식품, 식품과 영양소와의 관계, 우유의 성분, 노인영양에 대하여 잘 알고 있는 비율이 낮았으며, 인체의 영양소, 식단작성여부, 간식의 이론, 식품감별법에 대하여는 가장 낮은 비율을 나타냈다. 각 영양지식은 교육정도가 높을수록 영양지식이 높았고, 교육별 집단간의 유의한 차가 나타났다. (0.001

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High-efficiency development of herbicide-resistant transgenic lilies via an Agrobacterium-mediated transformation system (고효율의 아그로박테리움 형질전환법을 이용한 제초제저항성 나리 식물체 개발)

  • Jong Bo Kim
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.56-62
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    • 2023
  • Transgenic lilies have been obtained using Agrobacterium tumefaciens (AGL1) with the plant scale explants, followed by DL-phosphinothricin (PPT) selection. In this study, scales of lily plants cv. "red flame" were transformed with the pCAMBIA3301 vector containing the gus gene as a reporter and the blpR gene as a selectable marker, as well as a gene of interest showing herbicide tolerance, both driven by the CaMV 35S promoter. Using a 20-minute infection time and a 5-day cultivation period, factors that optimized and demonstrated a high transformation efficiency were achieved. With these conditions, approximately 22-27% efficiency was observed for Agrobacterium-mediated transformation in lilies. After transformation with Agrobacterium, scales of lilies were transferred to MS medium without selective agents for 2 weeks. They were then placed on selection MS medium containing 5 mg/L PPT for a month of further selection and then cultured for another 4-8 weeks with a 4-week subculture regime on the same selection medium. PPT-resistant scales with shoots were successfully rooted and regenerated into plantlets after transferring into hormone-free MS medium. Also, most survived putatively transformed plantlets indicated the presence of the blpR gene by PCR analysis and showed a blue color indicating expression of the gus gene. In conclusion, when 100 scales of lily cv. "red flame" are transformed with Agrobacterium, approximately 22-27 transgenic plantlets can be produced following an optimized protocol. Therefore, this protocol can contribute to the lily breeding program in the future.

Optimal supervised LSA method using selective feature dimension reduction (선택적 자질 차원 축소를 이용한 최적의 지도적 LSA 방법)

  • Kim, Jung-Ho;Kim, Myung-Kyu;Cha, Myung-Hoon;In, Joo-Ho;Chae, Soo-Hoan
    • Science of Emotion and Sensibility
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    • v.13 no.1
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    • pp.47-60
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    • 2010
  • Most of the researches about classification usually have used kNN(k-Nearest Neighbor), SVM(Support Vector Machine), which are known as learn-based model, and Bayesian classifier, NNA(Neural Network Algorithm), which are known as statistics-based methods. However, there are some limitations of space and time when classifying so many web pages in recent internet. Moreover, most studies of classification are using uni-gram feature representation which is not good to represent real meaning of words. In case of Korean web page classification, there are some problems because of korean words property that the words have multiple meanings(polysemy). For these reasons, LSA(Latent Semantic Analysis) is proposed to classify well in these environment(large data set and words' polysemy). LSA uses SVD(Singular Value Decomposition) which decomposes the original term-document matrix to three different matrices and reduces their dimension. From this SVD's work, it is possible to create new low-level semantic space for representing vectors, which can make classification efficient and analyze latent meaning of words or document(or web pages). Although LSA is good at classification, it has some drawbacks in classification. As SVD reduces dimensions of matrix and creates new semantic space, it doesn't consider which dimensions discriminate vectors well but it does consider which dimensions represent vectors well. It is a reason why LSA doesn't improve performance of classification as expectation. In this paper, we propose new LSA which selects optimal dimensions to discriminate and represent vectors well as minimizing drawbacks and improving performance. This method that we propose shows better and more stable performance than other LSAs' in low-dimension space. In addition, we derive more improvement in classification as creating and selecting features by reducing stopwords and weighting specific values to them statistically.

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PS-341-Induced Apoptosis is Related to JNK-Dependent Caspase 3 Activation and It is Negatively Regulated by PI3K/Akt-Mediated Inactivation of Glycogen Synthase Kinase-$3{\beta}$ in Lung Cancer Cells (폐암세포주에서 PS-341에 의한 아포프토시스에서 JNK와 GSK-$3{\beta}$의 역할 및 상호관련성)

  • Lee, Kyoung-Hee;Lee, Choon-Taek;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.57 no.5
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    • pp.449-460
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    • 2004
  • Background : PS-341 is a novel, highly selective and potent proteasome inhibitor, which showed cytotoxicity against some tumor cells. Its anti-tumor activity has been suggested to be associated with modulation of the expression of apoptosis-associated proteins, such as p53, $p21^{WAF/CIP1}$, $p27^{KIP1}$, NF-${\kappa}B$, Bax and Bcl-2. c-Jun N-terminal kinase (JNK) and glycogen synthase kinase-$3{\beta}$ (GSK-$3{\beta}$) are important modulators of apoptosis. However, their role in PS-341-induced apoptosis is unclear. This study was undertaken to elucidate the role of JNK and GSK-$3{\beta}$ in the PS-341-induced apoptosis in lung cancer cells. Method : NCI-H157 and A549 cells were used in the experiments. The cell viability was assayed using the MTT assay and apoptosis was evaluated by proteolysis of PARP. The JNK activity was measured by an in vitro immuno complex kinase assay and by phosphorylation of endogenous c-Jun. The protein expression was evaluated by Western blot analysis. Dominant negative JNK1 (DN-JNK1) and GSK-$3{\beta}$ were overexpressed using plasmid and adenovirus vectors, respectively. Result : PS-341 reduced the cell viability via apoptosis, activated JNK and increased the c-Jun expression. Blocking of the JNK activation by overexpression of DN-JNK1, or pretreatment with SP600125, suppressed the apoptosis induced by PS-341. The activation of caspase 3 was mediated by JNK activation. Blocking of the caspase 3 activation suppressed PS-341-induced apoptosis. PS-341 activated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, but its blockade enhanced the PS-341-induced cell death via apoptosis. GSK-$3{\beta}$ was inactivated by PS-341 via the PI3K/Akt pathway. Overexpression of constitutively active GSK-$3{\beta}$ enhanced PS-341-induced apoptosis; in contrast, this was suppressed by dominant negative GSK-$3{\beta}$ (DN-GSK-$3{\beta}$). Inactivation of GSK-$3{\beta}$ by pretreatment with lithium chloride or the overexpression of DN-GSK-$3{\beta}$ suppressed both the JNK activation and c-Jun up-regulation induced by PS-341. Conclusion : The JNK/caspase pathway is involved in PS-341-induced apoptosis, which is negatively regulated by the PI3K/Akt-mediated inactivation of GSK-$3{\beta}$ in lung cancer cells.

Effect of TNF-$\alpha$ Gene Transfer to Respiratory Cancer Cell Lines on Sensitivity to Anticancer drugs (호흡기계암세포주에서 TNF-$\alpha$ 유전자의 이입이 항암제 감수성에 미치는 효과)

  • Mo, Eun-Kyung;Lee, Jae-Ho;Lee, Kye-Young;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Choi, Hyung-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.3
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    • pp.302-313
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    • 1995
  • Background: Tumor necrosis factor(TNF) showed antitumor cytolytic effects on sensitive tumor cells in numerous in vivo and in vitro studies. But it could not be administered systemically to human because of severe systemic adverse effects at effective concentrations against tumor cells. Many studies showed that a high concentrations of TNF in the local milieu may evoke in vivo TNF-responsive mechanisms sufficient to suppress tumor growth. Recently developed technique of TNF gene transfer to tumor cells using retrovirus vector could be a good candidate for local TNF administration. TNF is also known to synergistically enhance in vitro cytotoxicity of chemotherapeutic drugs targeted to DNA topoisomerase II against TNF-sensitive tumor cell lines. In this study the in vitro chemosensitivity against DNA topoisomerase II targeted chemotherapeutic drugs was evaluated using some respiratory cancer cell lines to which TNF gene had been transferred. Method: NCI-H2058, a human mesothelioma cell line, A549, a human lung adenocarcinoma cell line and WEHI 164 cell line, a murine fibrosarcoma cell line were treated with etoposide and doxorubicin, which are typical topoisomerase II - targeted chemotherapeutic agents, at different concentration. The resultant cytotoxicity was measured by MIT assay. Then the cytotoxicity of the same chemotherapeutic agents was measured after TNF-$\alpha$ gene-transfer and the two results were compared. Results: The cytotoxicity was not increased significantly in WEHI164 cell line and A549 cell line but statistically significant increase was observed in H2058 cell line when TNF-$\alpha$ gene was transferred(p<0.05). Conclusion: These findings show that TNF-$\alpha$ gene transfer to respiratory cancer cell lines results in variable effects on chemosensitivity against topoisomerase II inhibitor among different cell lines in vitro and can be additively cytotoxic in certain selective tumor cell lines.

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