• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.347-354
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• 2007

Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in human health and the environment. Prior to GM plant release, applicants should provide the information on GM crops for approval. We carried out this study to provide the molecular data for risk assessment of the GM Chinese cabbage plants with insect-resistance gene, modified CryIAc, which we obtained by Agrobacterium-transformation. From the molecular analysis with GM Chinese cabbage, we confirmed the transgene copy number and stability, the expression of the transgene, and integration region sequences between the transgene and the Chinese cabbage genome. Based on the unique integration DNA sequences, we designed specific primer set to detect GM Chinese cabbage and set up the GM cabbage detection method by qualitative PCR analysis. Qualitative analysis with GM Chinese cabbage progenies analysis was revealed the same as the result of herbicide treatment. Our results provided the molecular data for risk assessment analysis of GM Chinese cabbage and demonstrated that the primer set proposed could be useful to detect GM Chinese cabbage.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.10a
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• pp.145-145
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• 2005

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2005.10a
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• pp.145-145
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• 2005

• Journal of The Korean Society of Grassland and Forage Science
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• v.20 no.2
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• pp.103-108
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• 2000

To produce of transgenic orchardgrass, the seed-derived calli of orchardgrass (Dactylis glomerata L.) co-cultivated with Agrobacterium turnefaciens EHAlOl harboring binary vector pIG121-Hm were selected with hygromycin and then transferred onto N6 regeneration medium containing 1 rngl l of NAA, 5 rngl l of kinetin, 250 rngl l of carbenicillin and 50 mg/ l of hygromycin. The efficiency of transformation was differed on cultivars, that is, 'Potomac' appeared 12% of transformation efficiency while 'Amba' did 5.5%. The addition of acetosyringone during co-cultivation was a key to successhl transformation of orchardgrass. Transgene fragments were identified by PCR analysis and the constitutive expression of GUS gene was confirmed by Northern blot analysis. (Key words : Acetosyringone, Agrobacterium tumefaciens, Orchardgrass (Dactylis glomerata L.), Transformation)

• Journal of Microbiology
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• v.43 no.2
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• pp.179-182
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• 2005

The inefficiency of in vivo gene transfer using currently available vectors reflects a major hurdle in cancer gene therapy. Both viral and non-viral approaches that improve gene transfer efficiency have been described, but suffer from a number of limitations. Herein, a fiber-modified adenovirus, carrying the small peptide ligand on the capsid, was tested for the delivery of a transgene to cancer cells. The fiber-modified adenovirus was able to mediate the entry and expression of a $\beta$-galactosidase into cancer cells with increased efficiency compared to the unmodified adenovirus. Particularly, the gene transfer efficiency was improved up to 5 times in OVCAR3 cells, an ovarian cancer cell line. Such transduction systems hold promise for delivering genes to transferrin receptor overexpressing cancer cells, and could be used for future cancer gene therapy.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.143-148
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• 2005

The coat protein mediated resistance to potato virus Y is assessed here in transgenic potato plants (Solanum tuberosum L., cv Claustar). Therefore, the corresponding cDNA from tunisian isolate of the virus was cloned into Agrobacterium tumefaciens binary vector. The transgenic lines were subsequently analysed for the presence and expression of the transgene. The CP cDNA copy number was determined for kanamycin resistant plants. Three selected transgenic lines and their S1 progeny resulting from tuber germination showed a high protection level against the virus. These data appear to support the hypothesis that the virus resistance is mediated by the translated viral coat protein expressed in transgenic potato lines.

• Korean Journal of Animal Reproduction
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• v.18 no.3
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• pp.175-182
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• 1994

Rat $\beta$-casein 유전자와 사람 성장호르몬 (hGH) 유전자의 융합유전자를 생쥐 수정란의 웅성전핵에 미세주입하여 형질전환생쥐 (transgenic mouse) 6계통을 확립하였다. 이들로부터 사람성장호르몬(hGH) 유전자의 발현 여부를 조사한 결과, 6계통중 4계통의 생쥐 유즙에서 hGH가 2~900 ng/ml 수준으로 발현되고 있었으며, 혈중에서는 hGH가 검출되지 않았다. 따라서 이들 형질전환생쥐에서 사람성장호르몬이 우선특이적으로 발현, 분비되고 있음을 알 수 있었다. 한편, 사람성장호르몬의 발현이 확인된 두 계통 (ChGH2-2, CChGH2)의 형질전환생쥐를 대상으로 하여 세대별, 산차별로 유즙내 사람성장호르몬의 함량을 조사한 결과, 3세대에 걸쳐, 또한 제1세대에서의 세차례 반복된 비유기에도 유즙내 사람성장호르몬은 지속적으로 분비되고 있었다. 이상의 결과는 형질전환생쥐에서의 외래유전자의 발현성은 반복되는 비유기와 여러 세대에 걸쳐 안정적으로 유지됨을 보여 주고 있다.

• Korean Journal of Breeding Science
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• v.40 no.2
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• pp.128-135
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• 2008

Herbicide resistance is the most common trait being tested and thus herbicide?resistant genetically modified plants are now the most widely cultivated worldwide. Here we developed herbicide?resistant transgenic Agrostis mongolica Roshev. by employing an efficient Agrobacterium?mediated transformation procedure with 25.2% of transformation efficiency. The identification and employment of regenerable and reproducible type of callus was one of the most critical factors to ensure success in this study. PCR analysis confirmed that the bar transgene was integrated into the genome of transgenic plants. The expression of 35S?bar gene was confirmed by Northern blot analysis. The transgenic plants showed complete resistance to herbicide, indicating that the bar gene is functional in transgenic plants.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.3
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• pp.358-363
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• 2008

The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

• Journal of Plant Biotechnology
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• v.47 no.2
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• pp.172-178
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• 2020

Fruit-specific promoters play an important role in the improvement of traits, such as fruit quality through genetic engineering. In tomato, the development of fruit-specific promoters was previously reported, but less attention has been paid to the promoters involved in the fruit development stage. In this study, we characterized the gene expression patterns of tomato alcohol dehydrogenase 2 (SlAdh2) in various tissues of wild-type tomato (cv. Ailsa Craig). Our findings revealed that SlAdh2 expression levels were higher in the developing fruit than in the leaves, stems, and flowers. The ProSlAdh2 region, which is expressed at different stages of fruit development, was isolated from tomato genomic DNA. Following this, it was fused with a β-glucuronidase reporter gene (GUS) and introduced into wild-type tomato using Agrobacterium-mediated transformation to evaluate promoter activity in the various tissues of transgenic tomato. The ProSlAdh2:GUS promoter exhibited strong activity in the fruit and weak activity in the stems, but displayed undetectable activity in the leaves and flowers. Interestingly, the promoter was active from the appearance of the green fruit (1 cm in size) to the well-ripened stage in transgenic tomatoes, indicating its suitability for transgene expression during fruit development and ripening. Thus, our findings suggest that ProSlAdh2 may serve as a potential fruit-specific promoter for genetic-based improvement of tomato fruit quality.