• Title/Summary/Keyword: Transgene

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Infertility of Transgenic Mice Experssing Human Growth Hormone Gene (사람 성장호르몬 유전자를 발현하는 형질전환생쥐의 불임성)

  • 한용만;강만종;이철상;유대열;이경광
    • Korean Journal of Animal Reproduction
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    • v.16 no.3
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    • pp.225-230
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    • 1992
  • Many transgenic mice expressing human growth hormone gene were infertile. To investigate the infertility of these transfenic mice, it was looked into the estrus cycle and sexual behaviour and also tested through in vitro fertilization whether the germ cells of these mice normal or not. The infertile female transgenic mice were mated to the fertile males of ICR strain, but in almost all of them the vaginal plugs were not detected and their estrus cycles by vaginal smear were almost irregular which kept up estrus or diestrus stage. Many male transgenic mice did not have the ability of sexual behaviour. Therefore the viability of germ cells in infertile male transgenic mice was investigated by in vitro fertilization, but the sperm were normally fertilized with the eggs and the transgene of parent was passed on to the progeny. These results consequently suggest that the infertility of transgenic mice experssing human growth hormone gene may be due to the physiological activity of human growth hormone, not germ cells.

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A Simple and Reliable Method for Preparation of Cross-Contamination-Free Plant Genomic DNA for PCR-Based Detection of Transgenes

  • Hwang, Seon-Kap;Kim, Young-Mi
    • BMB Reports
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    • v.33 no.6
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    • pp.537-540
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    • 2000
  • A simplified but reliable method was developed for the polymerase chain reaction (PCR)-based detection of genetically modified (GM) plants. The modified CTAB (mCTAB) method enabled us to prepare a high quality of genomic DNA from several hundred plant leaf samples in one day. Using DNA samples prepared from seven dicots and two monocots, approximately 1.75-kb regions spanning 17 S to 25 S ribosomal RNA genes were successfully amplified in a 2X PCR pre-mix containing BLOTTO. Further fidelity assessment of the mCTAB method by PCR analysis with Roundup Ready soybean (RRS) and non-RRS plants showed that the DNA samples prepared alternately from each of two lines were evidently free of cross-contamination. These results demonstrate that the mCTAB method is highly recommended for the rapid detection of transgenes in large numbers of leaf samples from diverse transgenic plants.

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Herbicide Resistant Turfgrass(Zoysia japonica cv. 'Zenith') Plants by Particle bombardment-mediated Transformation

  • Lim Sun-Hyung;Kang Byung-Chorl;Shin Hong-Kyun
    • Asian Journal of Turfgrass Science
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    • v.18 no.4
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    • pp.211-219
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    • 2004
  • Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

Use of Flp-Mediated Cassette Exchange in the Development of a CHO Cell Line Stably Producing Erythropoietin

  • Kim, Min-Soo;Lee, Gyun-Min
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1342-1351
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    • 2008
  • The feasibility of the use of Flp-mediated cassette exchange in the development of a CHO cell line, which produces erythropoietin (EPO) stably and largely, was investigated. A stable, high enhanced green fluorescence protein (EGFP)-producing clone was screened by extensive flow cytometric analysis. An EPO expression unit was targeted into the premarked locus of the stable parental clone by Flp-mediated cassette exchange and a correctly targeted clone (FC28T7) was obtained. The EPO production of FC28T7 was proven to be stable in long-term culture. Furthermore, the Flp-mediated cassette exchange did not alter the stable parental clone's characteristics concerning transgene expression level and stability. Taken together, the data obtained here indicated that the establishment of CHO cell lines stably producing a desired protein is achievable using Flp-mediated cassette exchange.

ASSESSMENT OF STABILITY AND ALLERGENICITY OF FOOD ALLERGENIC PROTEINS

  • Lee, J-H;Yoon, W-K;Han, S-B;Yun, S-O;Park, S-H;Lee, H-J;Yoon, P-S;Moon, J-S;Kim, H-C
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.180-180
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    • 2002
  • The potential allergenicity of the transgene products in genetically modified organisms (GMOs), has been an important issue. As a part of the risk assessment of GMOs, we investigated the physicochemical stability and the immunogenicity of food allergens to determine their allergenicity.(omitted)

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Resistance of SOD2-transgenic petunia line to oxidative stress

  • Lee, Su-Young;Han, Bong-Hee;Kim, Yeong-Tae;Kim, Jin-Seog
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.562-566
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    • 2010
  • SOD2-transgenic $T_3$ petunia line (A2-36-2-1-1-35) was treated with different levels of methyl viologen (MV) to determine its resistance to oxidative stress. Four (4) levels of MV (0, 100, 200, and $400\;{\mu}M$) were applied. The SOD2-transgenic $T_3$ petunia line exhibited a very significant oxidative stress resistance at the highest MV concentration ($400\;{\mu}M$) treatment compared to non-transgenic plant. RNA and protein expression of SOD2 transgene and higher parenchyma cell density in the transgenic petunias exhibiting resistance to oxidative stress proves its contribution to the expression of its resistance to oxidative stress.

Development of Tetracycline-regulated Adenovirus Expression Vector System

  • Son, Kyung-Hwa;Lee, Seung-Hoon;Kim, Jong-Sik;Choi, Jung-Joo;Lee, Je-Ho
    • Journal of Genetic Medicine
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    • v.3 no.1
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    • pp.33-37
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    • 1999
  • Recombinant adenovirus vector systems with strong promoters have been used to achieve high level production of recombinant protein. However, this overexpression system cause some problems such as disturbance of cell physiology and increment of cellular toxicity. Here, we showed a tetracycline-regulated adenovirus expression vector system. Our results showed that the expression level of transgene(p-53) was high and easily regulated by tetracycline. In addition, the maximal gene expression level of the tetracycline-controlled gene expression system was higher than that of the wild type CMV promoter system. Therefore, tetracycline-regulated adenoviral vector system could be applicable for regulatory high-level expression of toxic gene. Also, this system will be useful for functional studies and gene therapy.

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Herbicide-resistant Transgenic Mongolian Bentgrass (Agrostis mongolica Roshev.) obtained by Agrobacterium-mediated Transformation

  • Vanjildorj, Enkhchimeg;Bae, Tae-Woong;Song, In-Ja;Kim, Kyung-Moon;Lim, Yong-Pyo;Lee, Hyo-Yeon
    • 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.

Bombyx mori β-tubulin Promoter for High-level Expression of Heterologous Genes

  • Park, Kwanho;Goo, Tae-Won
    • International Journal of Industrial Entomology and Biomaterials
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    • v.39 no.1
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    • pp.22-28
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    • 2019
  • We previously isolated 9 clones that show stronger signal compared to Bombyx mori cytoplasmic actin gene (BmA3) by using a dot blot hybridization. In this study, we focused on one clone among these clones which has high amino acid similarity with ${\beta}$-tubulin gene of B. mori. This clone was ubiquitously expressed in all tissues and developmental stage of B. mori. As result of promoter assay using dual luciferase assay system, we found the highest transcription activity region (-750/-1) in the 5'-flanking region of ${\beta}$-tubulin gene, which has about 47 fold more intensive promoter activity than BmA3 promoter. Moreover, the ${\beta}$-tubulin promoter was normally regulated in Bm5, Sf9, and S2 cells. Therefore, we suggest that ${\beta}$-tubulin promoter may be used more powerful and effectively for transgene expression in various insects containing B. mori as a universal promoter.

Assessment of allergenicity of genetically modified foods (GMOs)

  • Lee, Jung-Hyun;Yoon, Won-Ki;Han, Sang-Bae;Yun, Si-On;Park, Sun-Hong;Lee, Hyun-Ju;Yoon, Pyung-Seop;Moon, Jae-Sun;Kim, Hyung-Chin;Kim, Hwan-Mook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.267.1-267.1
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    • 2002
  • The potential allergenicity of the transgene products in genetically modified organisms (GMOs). has been an important issue. As a part of the risk assessment of GMOs. we investigated the physicochemical stability and the immunogenicity of food allergens to determine their allergenicity. We have systematically evaluated the stability of food allergens in the gastrointestinal tract by using simple models of gastric (Stimulated gastric fluid) and intestinal (Stimulated intestinal fluid) digestion. (omitted)

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