• Title/Summary/Keyword: 식물 형질전환

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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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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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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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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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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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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.

The Role of S RNase Associated with Gametophytic Self-Incompatibility in Tomato (Lycopersicon peruvianum) (토마토 자가불화합성에 관여하는 S RNase 유전자의 기능)

  • 강나영;김명희;조규형;신동일;김달웅;박희성;정일경
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.219-226
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    • 2000
  • Lycopersicon peruvianum has a gametophytic self-incompatibility (GSI) mechanism controlled by a single genetic locus (S locus) with multiple alleles. S RNases, an allelic series of abundant stylar proteins, are products of the S locus in L. peruvianum and other Solanaceous plants. The $S_{11}$ RNase gene from L. peruvianum was introduced into a self-compatible (SC) species (Lycopersicon esculentum) to examine whether the expression pattern in the heterologous host mimics that in L. peruvianum. The resultant transgenic L. esculentum plants expressed the introduced gene highly in their styles, which is similar manner to the expresion in L. peruvianum. The $S_{11}$ RNase gene was expressed in the syle at a similar stage of flower development in both transgenic plants of L. esculentum and L. peruvianum without any morphological changes.

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Investigation of Transformation Efficiency of Rice Using Agrobacterium tumefaciens and High Transformation of GPAT (glycerol-3-phosphate acyltransferase) Gene Relative to Chilling Tolerance (Agrobacterium tumefaciens를 이용한 벼의 형질전환 효율의 검토 및 내한성 관련 GPAT (glycerol-3-phosphate acyltransferase) 유전자의 형질전환)

  • Seo, Mi-Suk;Bae, Chang-Hyu;Choi, Dae-Ock;Rhim, Seong-Lyul;Seo, Suk-Chul;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.85-92
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    • 2002
  • This study has been focused on improving transformation efficiency of rice using Agrobacterium tumefaciens. We have demonstrated the effect of this system when the GPAT gene related to the cold-resistance was transferred by Agrobacterium tumefaciens in rice. Transformation conditions were modified using intron $\beta$-glucuronidase (GUS) expression as a reporter gene in the rice. In this study, mature seed-derived calli of rice (Oruza sativa L. cv. Dongjin) were pre-cultured for 3 days and then infected with Agrobacterium. When this infected calli were cultured in the dark for 10 days on co-cu]lure medium containing 50 mg/L of CaCl$_2$, 30 mg/L of acetosyringone, 2 mg/L of 2,4-D, 120 mg/L of betaine, high GUS expression was observed. In the present transformation system, the efficiency of transformation of GPAT gene was about 54%. Stable integration of GPAT gene into chromosomal DNA was proven by southern blot analysis of genomic DNA isolated from T$_{0}$ progenies. The progenies (T1 generation) derived from primary transformant of 5 lines were segregated with a 3 (resistant) : 1 (sensitive ratio) in medium containing hygromycin. This high frequency transformation system can be used as a useful tool in transformation of another monocotyledon.n.

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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Organ Specific Expression of the nos-NPT II Gene in Transgenic Hybrid Poplar (형질 전환된 포플러에 대한 nos-NPT II 유전자의 기관별 발현 특성)

  • Chun, Young Woo;Klopfenstein, Ned B.
    • Journal of Korean Society of Forest Science
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    • v.84 no.1
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    • pp.77-86
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    • 1995
  • To effectively modify tree function with genetic engineering, transgenes must be expressed at the proper level in the appropriate tissues at suitable developmental stages. Toward understanding the spatial and temporal expression of transgenes in woody plants, transgene expression was evaluated in three greenhouse-grown, transgenic lines of Populus alba ${\times}$ P. grandidentata hybrid clone 'Hansen'. All transgenic poplar lines possess constructs containing the bacterial nopaline synthase(nos) promoter linked to a neomycin phosphotransferase II(NPT II) selectable marker gene. In addition, each transgenic poplar line contains one of the following gene constructs : 1) a wound-inducible potato proteinase inhibitor II (pin2) promoter linked to a chloramphenicol acetyltransferase(CAT) reporter gene. 2) a nos promoter linked to a PIN2 structural gene : or 3) a Cauliflower Mosaic Virus 35s promoter linked to a PIN2 structural gene. Polymerase chain reaction(PCR) was used to verify the presence of foreign genes in the poplar genome. Enzyme-linked immunosorbent assays(ELISAs) were used to evaluate organ specific expression of the nos-NPT II construct. NPT II expression was detected in leaves, petioles, stems, and roots of transgenic poplar, thereby indicating that the nos promoter is potentially effective for general constitutive expression of transgenes. NPT expression varied among transgenic poplar lines and among organs for one transgenic line, Tr15. With Tr15, NPT II levels were highest in older leaves and petioles. These results indicate that screening of several transgenic lines may be required to identify lines with optimal transgene expression.

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