• Title/Summary/Keyword: Biolistic system

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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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Efficient Gene Introduction into Rice Callus by Using Particle Inflow Gun System (Particle Inflow Gun을 이용한 벼 캘러스 내의 효율적 유전자 도입)

  • Song, In-Ja;Bae, Chang-Hyu;Choi, Dae-Ock;Ryo Akashi;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.223-228
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    • 2002
  • We have developed a homemade particle inflow gun (PIG) system which has simple operation method, low price and high gene introduction efficiency into rice callus. Rice callus were inflowed with gold particles containing DNA of a plasmid, pIG121Hm, harboring intron GUS ($\beta$-glucuronidase) gene, NPTII gene and HPT gene. For optimal GUS transient expression, the effects of parameters on DNA delivery efficiency of the PIG system was investigated by scoring transient GUS expression. The highest number of blue spots was observed at 16 mM of spemidine and 1.5 M of calcium chloride, respectively. And the amount of gold particles required for the best GUD expression was 2 mg. Optimum GUS transient expression was observed at target distance of 12 cm and helium pressure of 3.5 bar (50 psi). Gene introduction efficiency of the PIG system was observed almost similar to that of the Biolistic Gun (Bio-Rad Company). Since PIG system is simple to operate and one doesn't need disposable accessaries, the PIG system can be easily applied to various replication experiments.

A Routine System for Generation of Fertile Transgenic Rice Plants Using Biolistic Method

  • Lee Soo-In;Kim Cha-Young;Lim Chae-Oh;Choi Young-Ju;Kim Ho-Il;Lee Sang-Yeol;Lee Sung-Ho
    • Journal of Plant Biotechnology
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    • v.5 no.3
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    • pp.163-168
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    • 2003
  • A routine system based on particle bombardment of embryogenic callus for recovery of fertile transgenic rice (Oryza sativa L.) plants was developed. Embryogenic callus was established within 2-3 months from calli derived from mature seeds of Korean rice cultivar, Nagdongbyeo. The callus was bombarded with the plasmid pRQ6 containing the $\beta$-glucuronidase gene (gusA) and hygromycin phosphotransferase gene (hph, conferring resistance to hygromycin B), both driven by CaMV 35S promoter. Placement of cells on an osmoticum-containing medium (0.2 M sorbitol and 0.2 M mannitol) 4 hrs prior to and 16 hrs after bombardment resulted in a statistically significant increase with 3.2-fold in transient expression frequency gusA. In five independent experiments, the average frequency of transformation showing GUS activities was $8.86\%$. A large number of morphologically normal, fertile transgenic rice plants were obtained. Integration of foreign gene into the genome of $R_0$ transgenic plants was confirmed by Southern blot analysis. GUS and HPT were detected in $R_1$ progeny and Mendelian segregation of these genes was observed in $R_1$ progeny.

Optimization of Cymbidium transformation system by the particle gun techniques (DNA 입자총에 의한 Cymbidium속 난의 형질전환 조건 검토)

  • Hong, Kyung-Ae;So, In-Sup;Lee, Ok-Young;Cheong, Choong-Duk;Riu, Key-Zung;U., Zang-Kual
    • Applied Biological Chemistry
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    • v.39 no.4
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    • pp.260-264
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    • 1996
  • Process of particle bombardment for efficient transformation of Cymbidium virescence rhizome microcross sections was investigated using Biolistic particle delivery system with pBI121 harboring the ${\beta}-glucuronidase$(GUS) and the neomycin phosphotransferaseII(nptII). The best result was obtained from the combination of $1.11{\;}{\mu}m$ tungsten particles coated with pBl121, $77.33kg/cm^2$ helium pressure, 6.35 mm gap distance, and 7.0 cm target distance. Transient expression of the reporter gene, GUS, bombarded into the rhizome microsections was observed by the histochemical assay. The marker gene, nptII, delivered by bombarding the tungsten particles coated with the plasmid DNA was identified in the transformed rhizome by polymerase chain reaction.

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New Drug Delivery System Based on a Laser-Induced Shockwave (레이저 유도 충격파를 이용한 첨단 약물전달시스템 개발)

  • Han, Tae-Hee;Lee, Hyun-Hee;Gojani, Ardian B.;Yoh, Jai-Ick
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.34 no.1
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    • pp.67-71
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    • 2010
  • Impingement of a high power laser pulse (above 1 GW/$cm^2$) on a metal foil causes its ablation, which is characterized by a rapid expulsion of matter and the initiation of a strong shock wave inside the solid metal. The shock propagates through the foil and reverberates on the rear side, causing its deformation and microparticle ejection, which were deposited on the foil prior to ablation. Based on this principle, we are developing a new drug delivery system - Biolistic gun. Current study is focused on the controllability, stability, efficiency of the system, and characterization of the penetration shapes in various conditions. We have tested the system by applying direct and confined ablation. Several different media combinations were used for confinement-BK7 glass, water, BK7 glass with water, and succulent jelly(ultrasono jelly, RHAPAPHRM). Biological tissue was replicated by a 3% gelatin solution. Present data shows that the confinement results in enhancement of penetration shape reached by 5 um cobalt microparticles. Based on the analysis of the experimental results we observe that the penetration shape of microparticles can be controlled by adjusting the thickness of confinement media.