• Title/Summary/Keyword: transgenic

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Extracellular Superoxide Dismutase (EC-SOD) Transgenic Mice: Possible Animal Model for Various Skin Changes

  • Kim, Sung-Hyun;Kim, Myoung-Ok;Lee, Sang-Gyu;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • v.30 no.4
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    • pp.229-234
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    • 2006
  • We have generated transgenic mice that expressed mouse extracellular superoxide dismutase (EC-SOD) in their skin. In particular, the expression plasmid DNA containing human keratin K14 promoter was used to direct the keratinocyte-specific transcription of the transgene. To compare intron-dependent and intron-independent gene expression, we constructed two vectors. The vector B, which contains the rabbit -globin intron 2, was not effective for mouse EC-SOD overexpression. The EC-SOD transcript was detected in the skin, as determined by Northern blot analysis. Furthermore, EC-SOD protein was detected in the skin tissue, as demonstrated by Western blot analysis. To evaluate the expression levels of EC-SOD in various tissues, we purified EC-SOD from the skin, lungs, brain, kidneys, livers, and spleen of transgenic mice and measured its activities. EC-SOD activities in the transgenic mice skin were approximately 7 fold higher than in wild-type mice. These results suggest that the mouse overexpressing vector not only induces keratinocyte-specific expression of EC-SOD, but also expresses successfully functional EC-SOD. Thus, these transgenic mice appeared to be useful for the expression of the EC-SOD gene and subsequent analysis of various skin changes, such as erythema, inflamation, photoaging, and skin tumors.

MdMADS2 - transgenic chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) showing the reduction of the days to flowering

  • Han, Bong-Hee;Lee, Su-Young;Choi, Seong-Youl
    • Journal of Plant Biotechnology
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    • v.36 no.4
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    • pp.366-372
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    • 2009
  • This study was conducted to develop new lines expressing the characteristic of early flowering by introducing MdMADS2 gene in chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) ‘Zinba'. Transformation of chrysanthemum was conducted by Agrobacterium tumefaciens LBA4404 harboring the binary vector containing MdMADS2 controlled by double CaMV 35S promoters. Ninety three shoots were regenerated from 1,463 leaf segment explants cultured on the first selection medium (MS basal salts + 1.0 mg/L BA + 0.5 mg/L IAA + 10 mg/L kanamycin + 400 mg/L cefotaxime, pH 5.8) after co-cultivation, and 20 out of the 93 shoots rooted on the second selection medium containing 20 mg/L kanamycin and 400 mg/L cefotaxime. Many escapes (98.6%) were removed on the selection stage for rooting. Nineteen lines were confirmed as transgenic plant with transgene by PCR analysis. Six transgenic plants flowered 2-11 days earlier than non-transgenic plant without big change of phenotype, and especially, 3 (Mo-7, Mo-11, Mo-17) out of 6 transgenic lines showed a significant reduction in days to flowering compared to non-transgenic plant. Introduction and expression of MdMADS2 gene in them were confirmed by Southern and real-time PCR analyses, respectively.

Disease Occurrence in Transgenic Rice Plant Transformed with Silbene Synthase Gene and Evaluation of Possible Horizontal Gene Transfer to Plant Pathogens

  • Yu, Sang-Mi;Jeong, Ui-Seon;Lee, Ha Kyung;Baek, So Hyeon;Kwon, Soon Jong;Lee, Yong Hoon
    • Research in Plant Disease
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    • v.20 no.3
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    • pp.189-195
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    • 2014
  • Genetic engineering is being used to enhance disease resistance and nutritional value of crops including rice plant. Considering the fast-growing agricultural biotechnology and rapidly increasing global area of transgenic crops, the risk evaluation on environment is necessary. In this study, we surveyed the difference of disease occurrence between transgenic rice variety, Iksan526 transformed with peanut stilbene synthase gene and non-transgenic rice varieties, Dongjin and Nampyeong in the field. Moreover, the possibility of gene transfer from transgenic rice to bacterial and fungal pathogens was investigated. The results of this study indicated that there was no significant difference in the occurrence and severity of the diseases between Iksan526 and Dongjin or Nampyeong. In addition, the results suggested that rice pathogen, such as Xanthomonas oryzae pv. oryzae, Rhizoctonia solani and Magnaporthe grisea did not take up stilbene synthase and bar genes under natural conditions. Moreover the transformed DNA was not transferred to the pathogens even in repetitive contacts.

Amelioration of Cognitive Dysfunction in APP/PS1 Double Transgenic Mice by Long-Term Treatment of 4-O-Methylhonokiol

  • Jung, Yu-Yeon;Lee, Young-Jung;Choi, Dong-Young;Hong, Jin Tae
    • Biomolecules & Therapeutics
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    • v.22 no.3
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    • pp.232-238
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    • 2014
  • Alzheimer's disease (AD) is the most common neurodegenerative disease without known ways to cure. A key neuropathologic manifestation of the disease is extracellular deposition of beta-amyloid peptide (Ab). Specific mechanisms underlying the development of the disease have not yet been fully understood. In this study, we investigated effects of 4-O-methylhonokiol on memory dysfunction in APP/PS1 double transgenic mice. 4-O-methylhonokiol (1 mg/kg for 3 month) significantly reduced deficit in learning and memory of the transgenic mice, as determined by the Morris water maze test and step-through passive avoidance test. Our biochemical analysis suggested that 4-O-methylhonokiol ameliorated $A{\beta}$ accumulation in the cortex and hippocampus via reduction in beta-site APP-cleaving enzyme 1 expression. In addition, 4-O-methylhonokiol attenuated lipid peroxidation and elevated glutathione peroxidase activity in the double transgenic mice brains. Thus, suppressive effects of 4-O-methylhonokiol on $A{\beta}$ generation and oxidative stress in the brains of transgenic mice may be responsible for the enhancement in cognitive function. These results suggest that the natural compound has potential to intervene memory deficit and progressive neurodegeneration in AD patients.

Effects on the Development of Plutella xylostella and Spodoptera litura after Feeding on Transgenic Cabbage Expressing Potato Proteinase Inhibitor II and Bar Genes

  • Lee, Yeon-Hee;Lee, Sang-Guei;Park, Beom-Seok;Lee, Young-Su;Jin, Yong-Moon;Kim, Ho-il;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.145-150
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    • 2004
  • Cabbage plants were transformed with the potato proteinase inhibitor II (PINII) gene, bar gene, and hpt gene using Agrobacterium. The expression of the PINII gene was driven by its own promoter which was wound-inducible. Ten transgenic plants were obtained from medium containing hygromycin as a selection antibiotic. The integration and expression of PINII and bar genes were confirmed by Southern and Northern hybridization. Growth and development of diamondback moths (Plutella xylostella) and tobacco cutworm (Spodoptera litura) larvae were examined on $T_1$ plants. The weight of the larvae and pupae of these two insects grown on transgenic plants was not different compared to those grown on wild type plants. However, the pupation and emergence rate of diamondback moths and tobacco cutworms fed on some transgenic plants was lower than on wild type plants. These results suggest that the PINII transgene under the control of a wound-induced promoter may be used for control of insects in transgenic cabbage through reduction of insect progeny number.

Expression of Indica rice OsBADH1 gene under salinity stress in transgenic tobacco

  • Hasthanasombut, Supaporn;Ntui, Valentine;Supaibulwatana, Kanyaratt;Mii, Masahiro;Nakamura, Ikuo
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.75-83
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    • 2010
  • Glycine betaine has been reported as an osmoprotectant compound conferring tolerance to salinity and osmotic stresses in plants. We previously found that the expression of betaine aldehyde dehydrogenase 1 gene (OsBADH1), encoding a key enzyme for glycine betaine biosynthesis pathway, showed close correlation with salt tolerance of rice. In this study, the expression of the OsBADH1 gene in transgenic tobacco was investigated in response to salt stress using a transgenic approach. Transgenic tobacco plants expressing the OsBADH1 gene were generated under the control of a promoter from the maize ubiquitin gene. Three homozygous lines of $T_2$ progenies with single transgene insert were chosen for gene expression analysis. RT-PCR and western blot analysis results indicated that the OsBADH1 gene was effectively expressed in transgenic tobacco leading to the accumulation of glycine betaine. Transgenic lines demonstrated normal seed germination and morphology, and normal growth rates of seedlings under salt stress conditions. These results suggest that the OsBADH1 gene could be an excellent candidate for producing plants with osmotic stress tolerance.

Regulation of γ-Aminobutyric Acid Production in Tobacco Plants by Expressing a Mutant Calmodulin Gene

  • Oh, Suk-Heung;Cha, Youn-Soo
    • Journal of Applied Biological Chemistry
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    • v.43 no.2
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    • pp.69-73
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    • 2000
  • In order to understand the biological role of calmodulin in plants, transgenic plants expressing a mutant calmodulin (VU-4, Iys to ile-115) have been analyzed. We found that tobacco plants expressing VU-4 calmodulin have approximately twofold higher $\gamma$-aminobutyric acid (GABA) levels than the control plants. Cell suspension cultures established from the stem explants of the transgenic tobacco seedlings also have higher levels of GABA than the control cell cultures. Specific activity of glutamate decarboxylase (GAD), which catalyzes the decarboxylation of glutamate to $CO_2$ and GABA, of the transgenic tobacco cell extracts was about twofold higher than the activity of the control cell extracts. Western-blot analysis showed that the GAD is highly expressed in the transgenic tobacco plants. GAD partially purified from tobacco cell extracts showed approximately threefold $Ca^{2+}$/calmodulin-dependent activation. These data suggest that GABA synthesis in the transgenic tobacco plants is elevated, possibly due to higher levels of the calmodulin-dependent GAD enzyme and/or as a result of enhanced activation due to increased levels of the foreign calmodulin.

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Production of Transgenic Animals by the Testis-Mediated Gene Transfer I. Production of Transgenic Rats and Mice (정소실질내 유전자 도입에 의한 형질저환동물의 생산 I. 형질전환 흰쥐와 생쥐의 생산)

  • 윤창현;장규태;오석두;주학진;박미령;이병오
    • Korean Journal of Animal Reproduction
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    • v.22 no.2
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    • pp.145-152
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    • 1998
  • Many trials have been made to produce transgenic animals using sperm cells as a vector transferring foreign DNA into eggs, but reliable results are yet to be obtained (Brinster et al., 1989; Lavitrano et al., 1989; Bachiller et al., 1991; Sato et al., 1994). Recently, one of author(SO) demonstrated that mouse blastocysts derived from eggs fertilized by spermatozoa of male mice single injected with liposome-DNA complexes within the testis expressed thegene (Ogawa et al., 1995.) Here we report that a single injection of liposome-encapsulated DNAs into the testis of either male rats or mice resulted in successfully gene transfer to the postpartum progeny. The expression of mRNA derived from transgenes was also demonstrated in transgenic animals thus obtained. Further, the transmission of the exogenous gene to the descedants was confirmed in one line of transgenic rat up to F4 generation, indicating that the gene was stably incorporated into the germ line. Thus, direct single injection of foreign DNA into the testis provides a novel and convenient means to generate transgenic animals.

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Molecular Breeding of Transgenic Tomato Plants Expressing the ${\delta}-Endotoxin$ Gene of Bacillus thuringiensis subsp. tenebrionis (살충성 형질 전환 토마토 식물체의 분자 육종)

  • Rhim, Seong-Lyul
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.137-140
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    • 1998
  • The transgenic tomato plants showing the insecticidal activity against the coleopteran insect larvae have been bred to the 4th generation $(R_4)$. The Bacillus thuringiensis subsp. tenebrionis (B.t.t.)-toxin gene and the expression were detected in the $R_4$ transgenic plants. The expression of the toxin gene conferred a coleopteran insect larvae tolerance to the transgenic tomato plants. The ploidy levels of the $R_4$ transgenic plants were diploid. The results indicated that the toxin gene was inherrited to the next generation and expressed. Such a molecular breeding can provide a method for a permanent control of insects a agronomic relevance.

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Growth Performance and Field Evaluation of Herbicide-Resistant Transgenic Creeping Bentgrass

  • Lee, Ki-Won;Lee, Byung-Hyun;Seo, Bo-Ram;Kim, Jin-Seog;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.3
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    • pp.167-170
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    • 2013
  • This study examined the growth performance and field evaluation of the dual herbicide-resistant transgenic creeping bentgrass plants. The effect of glyphosate treatment on the herbicide resistance of the transgenic creeping bentgrass plants was determined, and the non-transgenic control plant withered at the concentration $11{\mu}g/mL$ or higher whereas the transgenic creeping bentgrass plants survived the treatment at the concentration of $3,000{\mu}g/mL$, and the increase of the plant length was repressed as the glyphosate treatment concentration was increased. At field evaluation, glufosinate-ammonium and glyphosate were simultaneously treated to investigate the weed control effect. The results showed that more than 90% of the weeds withered four week after herbicide treatment, while the transgenic creeping bentgrass plants continued to grow normally. Therefore, the dual herbicide-resistant creeping bentgrass plants may be able to greatly contribute to the efficiency of weed control and to the economic feasibility of mowing in places such as golf courses.