• Title/Summary/Keyword: Agrobacterium tumefaciens.

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Regeneration of Tobacco Tissue Introduced with the Maize Transposable Element Activator (옥수수 전이인자 Ac가 도입된 연초조직의 재분화)

  • 박성원;최광태;박지창;김영진
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.34-41
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    • 1991
  • To explore the possibility of introducing Zea mays transposable element Ac(activator) which can be used as a mutagen and gene tag in tobacco plants other than maitre, we tried to introduce a cloned Ac element into tobacco cells by an Agrobacterium tumefaciens binary vector system. Transformation of N. babacum cv. Burley 21 tissues and regeneration to whole plant were carried out. The frequency of the transformed callus induced in shoot induction media was higher than that of transformed callus induced in callus induction media. However, the calli were not grown in the second selection media, and became yellow senescent calli. Regenerated tobacco plantlets with foreign gene were also obtained in shoot induction media containing 100 $\mu\textrm{g}$/ml kanamycin and 100$\mu\textrm{g}$/ml carbenicillin. The leaf tissues of transformant was also resistant to 1000 $\mu\textrm{g}$/ml kanamycin. The chromosomal DNAs of transformant and normal plant of N. tabacum were digested by EcoR I and Hind III but not by Pst I.

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Double-Stranded RNA-Dependent Protein Kinase Gene Expression in Tobacco Plant (연초식물체에서의 dsRNA 의존성 인산화 효소 유전자 발현)

  • 이청호;박희성
    • Korean Journal Plant Pathology
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    • v.11 no.2
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    • pp.173-178
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    • 1995
  • 동물계에서 항바이러스와관련된 dsRNA 의존성 인산화 효소(PKR)의 유전자를 식물체에서 발현시킬 경우 PKR에 의한 단백질합성 및 식물바이러스의 증식조절 가능성에 대한 기초자료를 확보하기 위하여 사람에서 분리된 PKR cDNA를 Agrobacterium 방법에 의하여 연초식물체(Nicotiana tabacum cv. Xanthi-nc)로 형질전환시켰다. HindIII/PstI처리에 의해 얻어지는 약 1.8kb의 phPKR cDNA절편을 일련의 유전자 조작 방법을 통하여 식물발현벡터인 pBI121에 도입하여, p12168을 재조합하였다. 이를 A. tumefaciens LBA 4404에 형질전환시켜 연초식물체형질 전환에 이용하였다. 2mg/l BA와 0.5mg/l NAA가 포함되고 100$\mu\textrm{g}$/ml의 kanamycin이 첨가된 MS배지에서 shooting시킨 후 phytohormone이 첨가되지 않은 MS배지상에서 rooting을 시켜 형질전환 연초식물체를 얻었으며, 형질전환식물체는 정상식물체와 유사한 생육양상을 나타내었다. 형질전환식물체의 유전자도입은 hPKR cDNA의 전사부여는 RT-PCR 방법에 의하여 확인되었다.

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Cloning of CAB cDNA encoding chlorophyll a/b binding protein of photosystem II in Korean ginseng and Use in Plant (고려인삼 광계 II Chlorophyll a/b binding Protein 유전자(CAB)의 cloning 및 식물에의 활용연구)

  • 김갑식;이기원;이종철;여운형;채순용;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.21 no.2
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    • pp.152-159
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    • 1999
  • A CAB cDNA clone(pKGCAB) encoding the light harvesting chlorophyll a/b binding protein of the semi-shade plant, Korean ginseng(Panax ginseng C. A. Meyer) was isolated by the one-way path random sequencing of ginseng cDNA library clones and transgenic tobacco plants(Nicotiana tabacum NC82) were produced by the transformation of this ginseng CAB gene in use of Agrobacterium tumefaciens LBA4404. The CAB gene showed type 1 structure of LHCP-II, 84% similarity in nucleotide sequence and 92% in amino acid sequence to that of Nicotiana tabacum CAB40, respectively. Seed germination and initial growth of the transgenic tobacco plants transformed with the cDNA fragment were accelerated under low light intensity compared with those of normal tobacco plant, that may result from the higher light sensitivity of the transgenic plants than that of the normal.

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Expression of de novo Designed High Nutritional Peptide (HEAAE) in Tobacco

  • Kim, Jae-Ho;Lee, Chang-Kook;Hong, Bun-Shik
    • Journal of Microbiology and Biotechnology
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    • v.7 no.2
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    • pp.138-143
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    • 1997
  • We have designed and constructed a gene encoding novel high essential amino acid encoding protein(HEAAE). The resultant DNA fragment was tested for in vitro and in vivo expression and then cloned into plant expression vector pBI121, under the control of the cauliflower mosaic virus 35S promoter. Agrobacterium tumefaciens, strain LBA4404, was subsequently transformed with this new construct and Nicotiana tabacum var. Xanthi transgenic plants were obtained. DNA analysis by Southern procedure confirmed the presence of the multi-copy number of genes in the transformed plants. Analysis of RNA and protein synthesized in these transgenic plants demonstrated the stable expression of this gene.

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A target-specific bioassay for screening of bioactive AHL-analogues from natural products

  • Kim, Young-Hee;Kim, Jung-Sun;Park, Sung-Hoon
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.411-414
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    • 2002
  • Acylated homoserine lactones (AHLs) are membrane-permeant signal molecules responsible for biofilm formation of gram-negative bacteria via a unique mechanism known as quorum sensing. A target specific bioassay employing the AHL-responsive Agrobacterium tumefaciens reporter strain has been developed to identify new AHL-like compounds from natural products, which could be developed into antifouling compounds. By varying the X-gal concentration, incubation time, solvent for sample preparation and the sample loading procedure, it was possible to detect low level AHLs up to $10^1nM$. The length of the acyl chain of the AHLs was found to affect the sensitivity of this bioassay.

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Transgenic Tobacco Plants Introduced with cDNA of Cucumber Mosaic Virus Satellite RNA (오이 모자이크 바이러스 위성RNA의 cDNA가 도입된 형질전환 담배의 육성)

  • 이상용;홍은주;최장경
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.80-86
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    • 1995
  • The cDNA of CMV-As satellite RNA was introduced into tobacco plants (Nicotiana tabacum cv. Samsun NN) using a binary Ti plasmid vector system of Agrobacterium tumefaciens. The cDNA of satellite RNA introduced into tobacco plants was detected by polymerase chain reaction (PCR) and molecular hybridization analyses. Symptom development was distinctly suppressed in the transgenic tobacco plants when inoculated with CMV-Co. CMV concentration in the transgenic tobacco plants was decreased to 1/40 of non-transgenic tobacco plants. The kanamycin resistance gene of the transgenic tobacco plants was also detected in the progeny.

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PVY Resistant Transgenic Potato Plants (cv Claustar) Expressing the Viral Coat Protein

  • Gargouri-Bouzid Radhia;Jaoua Leila;Mansour Riadh Ben;Hathat Yemna;Ayadi Malika;Ellouz Radhouane
    • 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.