• Title/Summary/Keyword: Gene transformation

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Tumor targeted gene therapy (종양 표적 유전자 치료)

  • Kang, Joo-Hyun
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.5
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    • pp.237-242
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    • 2006
  • Knowledge of molecular mechanisms governing malignant transformation brings new opportunities for therapeutic intervention against cancer using novel approaches. One of them is gene therapy based on the transfer of genetic material to an organism with the aim of correcting a disease. The application of gene therapy to the cancer treatment has led to the development of new experimental approaches such as suicidal gene therapy, inhibition of oncogenes and restoration of tumor-suppressor genes. Suicidal gene therapy is based on the expression in tumor cells of a gene encoding an enzyme that converts a prodrug into a toxic product. Representative suicidal genes are Herpes simplex virus type 1 thymidine kinase (HSV1-tk) and cytosine deaminase (CD). Especially, physicians and scientists of nuclear medicine field take an interest In suicidal gene therapy because they can monitor the location and magnitude, and duration of expression of HSV1-tk and CD by PET scanner.

Production of bialaphos-resistant Nierembergia repens by electroporation

  • Shizukawa, Yoshiaki;Mii, Masahiro
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.219-226
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    • 2008
  • Transgenic plants with the herbicide-resistance gene (bar gene) were obtained via organogenesis from isolated mesophyll protoplasts of Nierembergia repens after applying electroporation. Transient ${\beta}-glucuronidase$ (GUS) activity of electroporated protoplasts assayed 2 days after applying an electric pulse showed that optimum condition (transient GUS activity 319 pmol 4 MU/mg per min and plating efficiency 2.43%) for electroporation was 0.5 kV/cm in field strength and $100{\mu}F$ in capacitance. The protoplasts electroporated with the bar gene at this condition initiated formation of microcolonies on medium after 2 weeks. After 4 weeks of culture, equal volume of fresh 1/2-strength Murashige and Skoog (MS) medium containing 0.2 mg/l bialaphos was added for selection of transformed colonies. After 6 weeks of culture, growing colonies were transferred onto regeneration medium containing 1.0 mg/l bialaphos, on which they formed adventitious shoots 1-2 months after electroporation. The adventitious shoots rooted easily after transfer onto MS medium with bialaphos lacking plant-growth regulators. Transformation of these regenerants with the bar gene was confirmed by Southern analysis. Some of the transformants showed strong resistance to the application of bialaphos solution at 10.0 mg/l.

Development Strategy for functional rice improved with human lactoferrin and enhancement of nutrient compounds (인체 모유 단백질 및 영양 성분 강화 고부가가치 기능성 쌀 생산 벼 품종 개발 전략)

  • Rhim Seong-Lyul;Lee Jin-Hyoung;Lee Hyo-Yeon;Suh Suk-Cheol
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2002.05a
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    • pp.48-50
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    • 2002
  • A strategy for development of a functional rice in proved with human lactoferrin and enhancement of nutrient compounds was planned. For the purposes we have cloned and characterized a human lactoferrin cDNA from human mammary gland cDNA library A endosperm storage vacuole targeting sequence and the cDNA fragment was linked to endosperm specific glutelin promoter. The fusion gene fragment was inserted into a binary vector containing MAR gene. In addition a new ${\beta}$-galactosidase gene from Bifidobacterium of human was used as a reporter gene in the vector system, Rice plants showing a high concentration of amino acids in the endosperm cells were developed by using a biochemical mutation and bred for the transformation with the binary vector system Finally we have established a transformation method for the rice endosperm cells.

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Induction of RNA-mediated Resistance to Papaya Ringspot Virus Type W

  • Krubphachaya, Pongrit;Juricek, Mila;Kertbundit, Sunee
    • BMB Reports
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    • v.40 no.3
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    • pp.404-411
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    • 2007
  • Transformation of cantaloupes with the coat protein (cp) gene of papaya ringspot virus type W (PRSV-W), Thai isolate, was used to introduce virus resistance. Binary vectors containing either the full length coat protein coding region under control of the 35S CaMV promoter(pSA1175), or the inverted-repeat of a coat protein coding region (pSA1304), were constructed and used for Agrobacteriummediated transformation of cotyledonary explants of the cantaloupe cultivar Sun Lady. Four independent transgenic lines were obtained using pSA1304 and one using pSA1175. Integration of the PRSV-W cp gene into the genome of these transgenic lines was verified by PCR amplification, GUS assays and Southern blot hybridization. In vitro inoculation of these lines with PRSV-W revealed that whereas the line containing pSA1175 remained sensitive, the four lines containing pSA1304 were resistant. The presence of small RNA species, presumably siRNA, corresponding to regions of the viral cp gene in transgenic lines resistant to PRSV-W supports the involvement of post-transcriptional gene silencing in the establishment of resistance.

Transformation and Expression of the PAT Gene in Arabidopsis Tryptophan Mutants

  • Lim, Seon-Hee;Kim, Young-Soon;Cheong, Hyeon-Sook
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.243-247
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    • 1996
  • Phosphoribosylanthranilate transferase (PAT) catalyzes the second step of the tryptophan biosynthetic pathway and is encoded by a single-copy gene that complements all the visible phenotypes of the tryptophan mutant (trp1-100) of Arabidopsis. The trp1-100 is blue fluorescent under UV light becuase it accumulates anthranilate. To obtain a plant with reduced PAT activity, PAT1 genes with several internal deletions in different promoter regions (pHS 101, pHS102, pHS104, pHS105, and pHS107) were induced into trp1-100 via Agrobacterium. Then, homozygous T3 plants were isolated and examined for blue fluorescence. Introduction of the PAT1 gene fusants results in the reversion of fluorescence phenotype except in the case of pHS105. These results prompted us to perform a parallel analysis of anthranilate synthase and PAT interms of the genetic complementation. A plant line carrying pHS105 gene fusant does not completely complement the blue fluorescence but it accumulates less anthranilate than trp1-100. The activity of PAT was reduced in the transgenic mutant as well. The plant carrying these constructs will add to the growing collection of molecular tools for the study of the indolic secondary metabolism.

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Prediction Model for the Cellular Immortalization and Transformation Potentials of Cell Substrates

  • Lee, Min-Su;Matthews Clayton A.;Chae Min-Ju;Choi, Jung-Yun;Sohn Yeo-Won;Kim, Min-Jung;Lee, Su-Jae;Park, Woong-Yang
    • Genomics & Informatics
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    • v.4 no.4
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    • pp.161-166
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    • 2006
  • The establishment of DNA microarray technology has enabled high-throughput analysis and molecular profiling of various types of cancers. By using the gene expression data from microarray analysis we are able to investigate diagnostic applications at the molecular level. The most important step in the application of microarray technology to cancer diagnostics is the selection of specific markers from gene expression profiles. In order to select markers of Immortalization and transformation we used c-myc and $H-ras^{V12}$ oncogene-transfected NIH3T3 cells as our model system. We have identified 8751 differentially expressed genes in the immortalization/transformation model by multivariate permutation F-test (95% confidence, FDR<0.01). Using the support vector machine algorithm, we selected 13 discriminative genes which could be used to predict immortalization and transformation with perfect accuracy. We assayed $H-ras^{V12}$-transfected 'transformed' cells to validate our immortalization/transformation dassification system. The selected molecular markers generated valuable additional information for tumor diagnosis, prognosis and therapy development.

Current status and outlook on genetic transformation of fruit trees in Korea (과수 형질전환 국내 연구 동향 및 전망)

  • Kim, Jeong-Hee;Song, Kwan-Jeong
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.408-413
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    • 2010
  • The paper reviewed research status and outlook of genetic transformation in fruit trees in Korea. Genetic transformation has been long considered as an alternative technique overcoming limitation of conventional breeding and conducted since early 1990's. An efficient genetic transformation was established with major cultivars of apple, even in Fuji and Gamhong, and some transgenic apple plants have been transferred into the greenhouse for further analysis of gene expression. A few successes of genetic transformation have been reported and application to a variety of cultivars tried in citrus and kiwifruit. Successful genetic transformation has not been reported in the other fruit trees including grapevine, yet and it is considered being at the level of research. Those factors including replacement of antibiotics as a selective agent, use of transgenic rootstocks and manipulation of gene expression at proper parts and developmental stages have been prerequisites for the rapid commercialization of transgenic fruit plants.

GUS gene expression and plant regeneration via co-culturing with Agrobacterium in grapevine (Vitis vinifera) (Agrobacterium 공동배양을 이용한 포도 재분화율 향상과 GUS 유전자의 발현)

  • Kim, Se-Hee;Kim, Jeong-Hee;Kim, Ki-Ok;Do, Gyeong-Ran;Shin, Il-Sheob;Cho, Kang-Hee;Hwang, Hae-Seong
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.308-314
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    • 2011
  • Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. In this study, methods for Agrobacterium tumefaciens-mediated transformation and plant regeneration of grapevine (Vitis vinifera) were evaluated. Tamnara, Heukgoosul, Heukbosek, Rizamat were co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, GUS gene as reporter gene and resistance to kanamycin as selective agent. Seven percent of the maximum regeneration frequency was obtained from co-cultivated with explants from Rizamat with LBA4404 strain on selection medium with kanamycin. The addition of acetosyringone, 200 ${\mu}m$ in virulence induction step was a key factor for successful GUS reporter gene expression in grapevine transformation. Transgenic plants showed resistance to kanamycin and the GUS positive response in leaf ($T_0$) stem ($T_0$) and petiole ($T_0$).