• Title/Summary/Keyword: 형질전환식물

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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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항산화효소 유전자 도입형질전환 담배의 수분스트레스에 대한 반응

  • 박용목;곽상수
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11b
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    • pp.231-232
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    • 2003
  • 환경스트레스에 대한 내성식물의 개발의 일환으로 항산화 유전자를 도입한 형질전환 식물체를 이용하여 수분스트레스에 대한 반응성을 비형질전환 식물체와 비교하였다. 심한 수분스트레스를 유도하였을 때, 비형질전환 식물체에 비해 형질전환 식물체가 저하하는 수분포텐셜에 대해 높은 기공전도도와 광합성 능력을 가진 것으로 나타났다.

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The simple assay of phosphinothricin acetyltransferase gene on the transgenic potato (형질전환 감자에서 제초제 저항성 유전자인 PAT gene의 간편한 확인)

  • 정재훈;양덕춘;방극수;최경화;한성수
    • Korean Journal of Plant Resources
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    • v.12 no.4
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    • pp.253-259
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    • 1999
  • In this study, three simple methods were established to confirm the transgenic potato plants. The leaf disc was used in the first method. After leaf discs of transgenic and non-transgenic potato were transfered into the liquid MS medium with bialaphos 5mg/l, 25 days, the chlorosis occurred in the non-transgenic leaf discs while it could not find in the transgenic leaf discs, In the second method, shoot tips of potato were transferred into MS medium supplemented with 0.5mg/l bialaphos and 0.6% agar. After 7-10 days, a lot of roots developed from the transgenic shoot tip, but the non-transgenic shoot tip was dead. The third method was using chlorophyll contents. Leaf discs were transferred into the liquid MS medium with bialaphos 0.5 mg/l. After 15 days, the content of chlorophyll A in transgenic plant was at least 2.5 times higher than in non-transgenic plant. In addition, the PAT enzyme activity were detected in the transgenic potato, but not detected in normal potato.

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Overexpression of the Metal Transport Protein1 gene (MTP1) in Arabidopsis Increased tolerance by expression site (금속전달 유전자(MTP1)의 과발현 애기장대에서 발현 위치에 따른 내성 증가 연구)

  • Kim, Donggiun
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.3
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    • pp.327-332
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    • 2019
  • Today's scientists try to remove heavy metals with many new technologies such as phytoremediation. One of the best cutting edge technologies is developing transgenic plants to remove certain heavy metal in soil. I constructed the transformation vector expressing T. goesingense Metal Transport Protein1 gene and TgMTP1: GFP genes. The transgenic plants were selected and confirmed the transformed genes into Arabidopsis thaliana genome. Expression was confirmed in several parts in Arabidopsis cells, tissues and organs. When TgMTP1 overexpressing Arabidopsis thaliana were subjected, transgenic plants showed higher heavy metal tolerance than non-transgenic. For further study I selected the transgenic plant lines with enhanced tolerance against four different heavy metals; Zn, Ni, Co, Cd. The accumulation of these metals in these plants was further analyzed. The TgMTP1 overexpressing Arabidopsis thaliana plant of selected lines are resistant against heavy metals. This plant is characterized by the expression of the MTP1 gene accumulating heavy metal in the vacuole and being simultaneously expressed on the plasma membrane. In conclusion, these plants may be used in plant purification applications, and as a plant with increased tolerance.

Expression of CAB (Chlorophyll a/b Binding Protein) Gene in Transformed Plants (CAB (Chlorophyll a/b Binding Protein) 유전자의 형질전환 식물체에서 발현)

  • 박성원;김선원;이영기;강신웅;이청호;이종철;최순용
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.41-45
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    • 2001
  • Transgenic tobacco plants were produced by the transformation of ginseng CAB gene using Agrobacterium tumefaciens LBA4404. The presence of CAB gene in the second generation of transgenic tobacco plant was confirmed by genomic PCR. The photosynthetic ability of transgenic plants was higher than normal tobacco plants and the maximum photosynthetic point of transgenic and normal tobacco plants was 500 $\mu$mol m$^{-2}$ s$^{-1}$ . The photosynthesis of C7, C11, 1, C14 cell lines was higher than normal plants at all the light intensities investigated. The photosynthesis of C2, C11, C14 cell lines in 90% dark condition was higher than normal plants. The chlorophyll contents of transgenic tobacco plants were almost same as normal plants. The % of dry weight, nicotine content, total sugar and nitrogen contents of harvested transgenic tobacco plant leaves were almost same as normal plants.

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Genetic Transformation of Panax ginseng with Herbicide Resistant Gene (제초제 저항성 유전자에 의한 인삼의 형질전환)

  • 양계진
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.353-357
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    • 2001
  • Transformation of ginseng plants was achieved by biolistic system with cotyledon explants and callus using phosphinothricin acetyl-transferase (PAT) gene resisting to a herbicide of Bialaphos. The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 355 promoter. The introduced NPT II and PAT genes of the transgenic ginseng plants were successfully identified by the PCR, and the survival test on the medium with basta. The transgenic ginseng plants were propagated using repetitive secondary embryogenesis. The transgenic ginseng plantlets had normal structures of roots and shoots, and dormant buds for new year sprouting. We transferred the transgenic plants to greenhouse and observed the continuing growth until a new year.

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Molecular Breeding of Transgenic Tomato Plants Expressing the ${\delta}-Endotoxin$ Gene of Bacillus thuringiensis subsp. tenebrionis (살충성 형질 전환 토마토 식물체의 분자 육종)

  • Rhim, Seong-Lyul
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.137-140
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    • 1998
  • The transgenic tomato plants showing the insecticidal activity against the coleopteran insect larvae have been bred to the 4th generation $(R_4)$. The Bacillus thuringiensis subsp. tenebrionis (B.t.t.)-toxin gene and the expression were detected in the $R_4$ transgenic plants. The expression of the toxin gene conferred a coleopteran insect larvae tolerance to the transgenic tomato plants. The ploidy levels of the $R_4$ transgenic plants were diploid. The results indicated that the toxin gene was inherrited to the next generation and expressed. Such a molecular breeding can provide a method for a permanent control of insects a agronomic relevance.

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Enhancement of Chloroplast Transformation Frequency by Using Mesophyll Cells Containing a Few Enlarged Chloroplasts from Nuclear Transformed Plants in Tobacco (적은 수의 거대 엽록체를 가진 핵 형질전환 식물체를 이용한 담배 엽록체 형질전환 빈도 제고)

  • Jeong, Won-Joong;Min, Sung-Ran;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.34 no.3
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    • pp.271-275
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    • 2007
  • In the chloroplast transformation process, a chloroplast containing transformed chloroplast genome copies should be selected over wild-type chloroplasts on selection medium. It is more effective for a cell to become homoplasmic if the cell contains smaller number of chloroplasts. Therefore, to reduce the number of chloroplasts in mesophyll cells in tobacco, we overexpressed FtsZ to generate transgenic plants, of which mesophyll cell contained a few enlarged chloroplasts contrast to a wild-type mesophyll cell containing approximately 100 chloroplasts. It was demonstrated that transgenic leaf tissues comprising cells with a few enlarged chloroplasts gave rise to approximately 40% higher frequency of chloroplast-transformed adventitious shoots.

Pathogene Resistance of cotton GST cDNA in Transgenic Scrophularia buergeriana Misrule (목화 Glutathione S-Transferase (GST) 유전자로 형질 전환된 현삼의 내병성 특성)

  • 강원희;임정대;이성호;유창연
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.297-304
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    • 2001
  • Scrophularia buergeriana Misrule has been contaminated with various pathogens in condition of field and storage period. This study was carried out for production of multiple stress resistance plant containing disease resistance that CGST gene expressed in transgenic Scrophularia buergeriana Misrule genome. Glutathione S-Transferases (GSTs) detoxify endobiotic and xenobiotic compounds by covalent linking of tripeptide glutathione to hydrophobic substrate. GST enzymes have been identified and characterized in insects, bacteria, and many plant species. A cDNA clone of GST was introduced into Scrophularia buergeriana Miquel by transformation with Agrobacterium tumefaciences. In coporation of the CGST gene into S. buergeriana Misrule was confirmed by PCR analysis of genomic DNA. Influence of exposure to darkness on the regeneration potential and transformation frequence were assessed. The activity of GST in transgenic plants was two times higher than that of non-transgenic plants. As a result of anti-microbe assays, the crude extract protein of transgenic plants showed the antimicrobial effects higher than control plants.

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Transgenic Plants Expressing an Antisense RNA of ALl-Gene from Tomato Golden Mosaic Virus(TGMV) (Tomato Golden Mosaic Virus(TGMV) AL1 -gene의 antisense RNA 발현 형질 전환 식물체)

  • 임성렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.147-152
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    • 1998
  • AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

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