• 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.

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.360-365
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• 2009

Over the last five years, we have conducted research on risk assessment of domestically developed genetically modified (GM) crops and found a number of factors which could delay risk assessment process. In this review, we described such cases and discussed the problem of transgene cassette integration, the lack of information on vectors, the poor quality control in seed production and absence of bioinformatic analysis on amino acid sequence homology before GM crop development. To solve these problems, we have suggested the introduction of the screening system of elite event before risk assessment process and quality control strategies for GM seed production. In addition, we suggested that the developers of GM crops should understand the importance of risk assessment and management for the commercialization of those crops and consider the biological and ecological characteristics of host plants. Consistent communications may need to be established between GM crop developers, risk assessors and risk managers at the initial stages of GM crop development to reduce trial-and-errors.

• 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.

• Journal of The Korean Society of Grassland and Forage Science
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• v.34 no.3
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• pp.187-192
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• 2014

Alfalfa (Medicago sativa L.) is one of the most important forage legumes in the world. It has been demanded to establish the efficient transformation system in commercial varieties of alfalfa for forage molecular breeding and production of varieties possessing new characteristics. To approach this, genetic transformation techniques have been developed and modified. This work was performed to establish conditions for effective transformation of commercial alfalfa cultivars, Xinjiang Daye, ABT405, Vernal, Wintergreen and Alfagraze. GUS gene was used as a transgene and cotyledon and hypocotyl as a source of explants. Transformation efficiencies differed from 0 to 7.9% among alfalfa cultivars. Highest transformation efficiencies were observed in the cultivar Xinjiang Daye. The integration and expression of the transgenes in the transformed alfalfa plants was confirmed by polymerase chain reaction (PCR) and histochemical GUS assay. These data demonstrate highly efficient Agrobacterium transformation of diverse alfalfa cultivars Xinjiang Daye, which enables routine production of transgenic alfalfa plants.

• 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.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2004.10a
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• pp.62-65
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• 2004

To protect the watermelon against soil-borne pathogens, we are currently producing disease-resistant transgenic root stock for the growth of watermelon, A defensin gene (J1-1) from Capsicum annum, a ACC deaminase gene from Pseudomonas syringae, a galactinol synthase (CsGolS) gene from Cucumis sativus, and a WRKY (CvWRKY2) gene from Citullus vulgaris were used as transgenes for disease resistance. The gene were transformed into a inbred line (6-2-2) of watermelon, Kong-dae watermelon and a inbred line (GO702S) of gourd, respectively, by Agrobacterium-mediated transformation. Putative transgenic plants were selected in medium containing 100mg/L kanamycin, and then integration of the genes into the genomic DNA were demonstrated by PCR analysis. Successful integration of the gene in regenerated plants was also confirmed by PCR (Figf 1), genomic Southern blot (Fig 2), RT-PCR (Fig 3), and Northern blot analysis(Fig 4). Several T1 lines having different transgene were produced, and disease resistance of the T1 lines are under estimation.

• 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.

• Plant Biotechnology Reports
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• v.2 no.4
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• pp.227-231
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• 2008

D-class cyclins play important roles in controlling the cell cycle in development and in response to external signals by forming the regulatory subunit of cyclin-dependent kinase (CDK) complexes. To evaluate the effects of D-class cyclins in transgenic rice plants, Arabidopsis cyclin D2 gene (CycD2) was linked to the maize ubiquitin1 promoter (Ubi1) and introduced into rice by the Agrobacterium-mediated transformation method. Genomic deoxyribonucleic acid (DNA), ribonucleic acid (RNA), and Western blot hybridizations of the Ubi1:-CycD2 plants revealed copy number of transgene and its increased expression in leaf and callus cells at messenger RNA (mRNA) and/or protein levels. The H1 kinase assay using the immunoprecipitates of protein extracts from the Ubi1:CycD2 plants and nontransgenic controls demonstrated that the introduced Arabidopsis CycD2 forms a functional CycD2/CDK complex with an unidentified CDK of rice. Shoot and root growth was enhanced in the Ubi1:CycD2 seedlings compared with nontransgenic controls, together, suggesting that Arabidopsis cyclin D2 interacts with a rice cyclin-dependent kinase, consequently enhancing seedling growth.

• Korean Journal of Animal Reproduction
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• v.22 no.2
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• pp.127-136
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• 1998

A chimeric gene comprising murine whey acidic protein(mWAP) and human growth hormone(hGH) was used to produce transgenic rats express hGH and secrete it into the blood. Two lines of transgenic rats carrying the mWAP/hGH construct were established; High line was characterized by relatively high levels of serum hGH, and low line had relatively low levels. The secretory profiles of rat GH(rGH) as well as hGH, the transgene product, were obtained in transgenic males and females of low line; both hGH and rGH serum levels were flattened with no episodic fluctuations, and the overall mean concentration of rGH was significantly lower than in normal littermates. Although the animals of High line showed an acceles, as assessed by vaginal opening and occurrence of first ovulation, advanced by 7∼8 days in both lines of animals. Accordingly, the body weight at puberty of low line transgenic females was much lower than that of normal littermates, indicating that continuous hGH expression could induce precocious puberty without enhancing the growth rate.

• Horticultural Science & Technology
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• v.32 no.3
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• pp.375-381
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• 2014

Transgenic potato was generated to express recombinant 5 repeated ${\beta}$-amyloid ($A{\beta}$) peptides, potential antigens to be applied as a preventive accine for Alzheimer's disease using Agrobacterium mediated transformation. The $A{\beta}$ peptides were fused to the human IgG Fc fragment enhancing protein and KDEL, which is the endoplasmic reticulum (ER) retention signal ($5A{\beta}$-FcK). The $5A{\beta}$-FcK, was expressed under the control of the duplicated 35S promoter. PCR analysis confirmed the presence of the transgene in several transgenic potato lines. Southern blot analysis showed only a single gene copy number in transgenic line 22, whereas multiple gene copy numbers were shown for transgenic lines 31 and 44. Northern blot analysis showed that line 22 had stronger mRNA levels when compared to lines 31 and 44. Immunoblot analysis confirmed that the $5A{\beta}$-FcK protein was expressed in the transgenic potato plant. These results indicate that $5A{\beta}$ fused to Fc can be expressed in potato plants.