• Title/Summary/Keyword: Particle Bombardment

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Methods for Introduction of the Atmospheric Nitrogen Fixing Ability to Plants

  • PreiningerE;BokaK;ZatykoJ;KoranyiP;GyurjanI
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.31-38
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    • 1999
  • An artificial symbiosis was established between diazotropic Azomonas insignis and strawberry (Fragaria x ananassa). The partnership was created by in vitro techniques through callus induction and organogenesis. The basis of this partnerships is the bacterial dependence on the plants metabolic activity, using maltose in the medium as a carbon and energy source which can be utilized by the plant cells only. The presence of bacteria in the intercellular spaces of the callus tissues and regenerated plants was proven by microscopic techniques. Nitrogenase activity could also be detected in the plant tissues. For successful and high frequency introduction of bacteria to the plant tissues, biolistic gun method was used. On the basis of the DNA transfer method, Azotobacter vinelandii bacteria were delivered directly into strawberry tissues by the particle bombardment. This was the first use of living bacteria as microprojectils for bombardment of plant tissues. The treatment was successful, the presence of bacteria in the developing callus tissue and regenerated plants were detected by light and electron microscopy.

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Effect of cultivar and ascorbic acid on in vitro shoot regeneration and development of bombardment-mediated plastid transformation of tomato (Lycopersicon esculentum) (토마토 재분화 효율 향상 및 엽록체 형질전환 조건)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Jong-Bum;Lee, Seung-Bum;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.77-83
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    • 2010
  • Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

Plastid Transformation of Soybean Suspension Cultures

  • Zhang, Xing-Hai;Archie R.Portis. Jr.;Jack M.Widholm
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.39-44
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    • 2001
  • Plastid transformation was attempted with soybean [Glycine max (L.) Merr.] leaves and photoautotrophic and embryogenic cultures by particle bombardment using the transforming vector pZVII that carries the coding sequences for both subunits of Chlamydomonas reinhardtii Rubisco and a spectinomycin resistance gene (aadA). Spectinomycin resistant calli were selected from the bombarded leaves but the transgene was not present, indicating that the resistance was due to mutations. The Chlamydomonas rbcL and rbcS genes were shown to be site-specifically integrated into the plastid genome of the embryogenic cells with a very low transformation efficiency. None of the transformed embryogenic lines survived the plant regeneration process so no whole plants were recovered. This result does indicate that it should be possible to insert genes into the plastid genome of the important crop soybean if the overall methods are improved.

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Development of transgenic sweet potato producing human lactoferrin (인체 락토페린 생산 형질전환 고구마 개발)

  • Min, Sung-Ran;Kim, Jae-Wha;Jeong, Won-Joong;Lee, Young-Bok;Liu, Jang R.
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.224-229
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    • 2009
  • Human lactoferrin is an iron-binding glycoprotein with many biological activities, including the protection against microbial and virus infection and stimulation of the immune system. We introduced a human lactoferrin (hLf) cDNA under the control of 35S promoter into sweet potato by particle bombardment. Transgenic plants were regenerated via somatic embryogenesis. Transgenic plants were produced typical tuberous roots in soil. PCR, Southern and northern analyses confirmed that the hLf cDNA was incorporated into the plant genome and was properly expressed in plants. Western blot analysis showed that the 80 kDa full length hLf protein was produced in transgenic tuberous roots. Overall results indicated that sweet potato would be an excellent host to produce human therapeutic proteins.

Discharge Characteristics from a Water-Penpoint-to - Insulator-Paper-Covered-Plate Airgap (수성(水性)펜침(針) 대(對) 절연지부착평판간(絶緣紙附着平板間)의 방전특성(放電特性))

  • Moon, Jae-Duk;Lee, Dae-Hee
    • Proceedings of the KIEE Conference
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    • 1994.07b
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    • pp.1586-1588
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    • 1994
  • Some interesting corona characteristics of a airgap of penpoint-to-plate with a tracing insulator paper has been investigated in a temperature and humidity controlled metal chamber. It is found that the positive and the negative carriers in the plasma region could be understood from the waterdrop traces on the paper, which indicate the bombardment by the one of electron or positive gas particle from the plasma region of near the point. It is also found that a corona discharge could be used as a means of a humidifying, since the discharge makes dispersion of watersprays from the penpoint. And the negative corona was more effective for waterspray dispersion than the positive ones.

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Plasma Treatment Effects on Tungsten Oxide Hole Injection Layer for Application to Inverted Top-Emitting Organic Light-Emitting Diodes

  • Kim, Joo-Hyung;Lee, You-Jong;Jang, Yun-Sung;Kim, Doo-Hyun;Hong, Mun-Pyo
    • 한국정보디스플레이학회:학술대회논문집
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    • 2009.10a
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    • pp.354-355
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    • 2009
  • In the fabrication of inverted top-emitting organic light emitting diodes (ITOLEDs), the sputtering process is needed for deposition of transparent conducting oxide (TCO) as top anode. Energetic particle bombardment, however, changes the physical properties of underlying layers. In this study, we examined plasma process effects on tungsten oxide ($WO_3$) hole injection layer (HIL). From our results, we suggest the theoretical mechanism to explain the correlation between the physical property changes caused by plasma process on $WO_3$ HIL and degradation of device performances.

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Transformation of the Diatom Phaeodactylum tricornutum with its Endogenous (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase Gene (Phaeodactylum tricornutum의 (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase 유전자의 형질전환)

  • Shin, Bok-Kyu;Jung, Yu-Jin;Kim, Sang-Min;Pan, Cheol-Ho
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.273-279
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    • 2015
  • Phaeodactylum tricornutum is a model diatom that its genomic information and biological tools are well established. In this study, a gene encoding (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (PtHDR), a terminal enzyme of the methylerythritol phosphate pathway regulating chlorophyll and carotenoid biosynthesis, was isolated from P. tricornutum. The isolated gene was cloned into pPha-T1 vector containing fcpA promoter to prepare pPha-T1-HDR plasmid. As a positive control, pPha-T1-eGFP plasmid was constructed with egfp gene. Stable nuclear transformation was carried out with these plasmids by particle bombardment method and zeocin resistant colonies of P. tricornutum were selected on f/2 agar plate. In result, transformation efficiency was evaluated according to the amount of plasmid DNA coated with gold particles. Integration of introduced plasmids was confirmed with genomic DNA of each transformant by polymerase chain reaction. The eGFP fluorescence was visible in the cytoplasm, indicating that eGFP was successively expressed in P. tricornutum system. The transcript level of exogenous Pthdr gene was evaluated with the obtained transformants. The results presented here demonstrated that introduction of Pthdr gene into P. tricornutum chromosome succeeded and expression of PtHDR was enhanced under the fcpA promoter.

Driving Characteristics of Flexible Reflective Display Using Carbon Nanotube Electrode (탄소나노튜브 전극을 이용한 플렉시블 반사형 디스플레이의 구동 특성)

  • Hwang, In-Sung;Kim, Young-Cho
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.25 no.6
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    • pp.451-455
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    • 2012
  • To compare an electrical and optical characteristics of indium tin oxide (ITO) and carbon nanotube (CNT) electrode on flexible and reflective display, we fabricate two charged particle-type display panels under the same panel condition of which the width of ribs is 10 ${\mu}m$, the cell size is $300{\mu}m{\times}300{\mu}m$, the q/m value of the white particles is -4.3 ${\mu}C/g$ and that for the black is +1.3 ${\mu}C/g$, and the cell gap is 75 ${\mu}m$, 125 ${\mu}m$, and 175 ${\mu}m$. We use plastic substrates coated with ITO and CNT electrode. To evaluate optical property, we measure a response time of particles using a laser and a photodiode. Threshold and driving voltages of CNT electrode according to the sheet resistance of 300, 600, 1,000 (ohm/sq) are compared with ITO electrode of 10 (ohm/sq). A response time of the CNT panel is similar to that of ITO panel, but the threshold and driving voltages of CNT panel are higher than that of ITO panel, inducing a large bombardment of the particles and shortening the lifetime of the panel. High difference of a threshold and a driving voltage of CNT panel will induce an particle clumping, resulting degradation of the panel. A bending radius of the fabricated CNT panel is 18 ${\mu}m$.

Genetic Transformation of Chrysanthemum with Cold Regulated Gene (BN115) (저온저항성 유전자를 이용한 국화 형질전환)

  • Han, Soo-Gon;Choi, In-Young;Kang, Chan-Ho;Ko, Bok-Rai;Choi, Joung-Sik;Lee, Wang-Hyu
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.19-25
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    • 2006
  • With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

Analysis of Upstream Regulatory Region from Populus nigra × P. maximowiczii by Inverse PCR Technique (Inverse PCR 기법(技法)을 이용(利用)한 양황철 DNA의 Regulatory Region의 탐색(探索))

  • Son, Suk Gyu;Hyun, Jung Oh
    • Journal of Korean Society of Forest Science
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    • v.87 no.3
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    • pp.334-340
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    • 1998
  • This research was conducted to identify plant regulatory regions by gene tagging method. A promoterless GUS coding sequence was introduced to Populus nigra ${\times}$ P. maximowiczii via Agrobacterium strains(LBA4404/EHA101), and putative transgenic poplars were selected by culturing on medium containing G418($60mg/{\ell}$) and by GUS assay. Among them one positive plant was to amplify the native sequences flanking to the introduced GUS gene in plant genome by inverse PCR method and from this 730 by DNA product was obtained. After subcloning and sequencing, it has 88% homology to the Eucalyptus gunnii CAD(cinnamyl alcohol dehydrogenase) gene. The GUS gene fused with the putative promoter reinserted into poplar leaves by particle bombardment method to test the funtional promoter activity. Upon staining with X-gluc, many blue spots appeared on the leaf segments bombarded by the chimeric gene 2-3 days, thus the isolated DNA fragment contain some possible coding region as well as a putative regulatory sequences of poplar CAD gene.

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