• Title/Summary/Keyword: transient expression system

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Impact of Physical, Chemical and Biological Factors on Lily (Lilium longiflorum cv. Georgia) Pollen Growth and GUS Expression Via Agro-infiltration (물리적, 화학적, 생물적 요인에 의한 백합 (Lilium longiflorum cv. Georgia) 화분의 생장 및 Agro-Infiltration을 이용한 GUS 발현)

  • Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.279-283
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    • 2004
  • To lily (Lilium longflorum cv. Georgia) pollen, impacts by some physical, chemical and biological factors were examined in respects of its growth and transient gene expression via agro-infiltration. Rolling movement in liquid medium or vacuum pressure during Agro-infiltration was regarded as a impact that should be minimized for normal pollen growth. Pollen growth was maintained well in relatively broad range of temperature (19 to 27$^{\circ}C$) or pH (5.0 to 8.0). Chemical factors such as cefotaxime (up to 300mg/L), acetosyringone (up to 800 $\mu$M) and syringealdehyde (up to 800 $\mu$M) did not show any harmful effects but kanamycin severely did even at concentration as low as 25mg/L in some cases. For GUS gene expression, acetosyringone at 200 to 400 $\mu$M slightly improved the efficiency while syringealdehyde did not. Brief agro-infiltration followed by 18 hr of co-incubation of pollen along with Agrobacterium was suggested as a condition basically required for the transient expression system using lily pollen regardless of the presence of acetosyringone.

Two-color Transient Grating Spectroscopy of a Two-level System

  • Kwak, Kyoung-Won;Cho, Min-Haeng;Fleming, Graham R.;Agarwal, Ritesh;Prall, Bradley S.
    • Bulletin of the Korean Chemical Society
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    • v.24 no.8
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    • pp.1069-1074
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    • 2003
  • A theoretical description and experimental demonstration of homodyne-detected two-color transient grating (2-C TG) signal are presented. By treating the coupled bath degrees of freedom as a collection of harmonic oscillators and using a short-time expansion method, approximated nonlinear response functions were obtained. An analytic expression for the two-color transient grating signal was obtained by carrying out relevant Gaussian integrals. The initial rising and decaying parts of the 2-C TG signal is shown to be critically dependent on the ultrafast inertial component of the solvation correlation function. The experimental results confirm the predictions of the theoretical model.

Functional characterization of ABA signaling components using transient gene expression in rice protoplasts

  • Song, In-Sik;Moon, Seok-Jun;Kim, Jin-Ae;Yoon, Insun;Kwon, Taek-Ryoun;Kim, Beom-Gi
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.109-109
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    • 2017
  • The core components of ABA-dependent gene expression signaling have been identified in Arabidopsis and rice. This signaling pathway consists of four major components; group A OsbZIPs, SAPKs, subclass A OsPP2Cs and OsPYL/RCARs in rice. These might be able to make thousands of combinations through interaction networks resulting in diverse signaling responses. We tried to characterize those gene functions using transient gene expression for rice protoplasts (TGERP) because it is instantaneous and convenient system. Firstly, in order to monitor the ABA signaling output, we developed reporter system named pRab16A-fLUC which consists of Rab16A promoter of rice and luciferase gene. It responses more rapidly and sensitively to ABA than pABRC3-fLUC that consists of ABRC3 of HVA1 promoter in TGERP. We screened the reporter responses for over-expression of each signaling components from group A OsbZIPs to OsPYL/RCARs with or without ABA in TGERP. OsbZIP46 induced reporter most strongly among OsbZIPs tested in the presence of ABA. SAPKs could activate the OsbZIP46 even in the ABA independence. Subclass A OsPP2C6 and -8 almost completely inhibited the OsbZIP46 activity in the different degree through the SAPK9. Lastly, OsPYL/RCAR2 and -5 rescued the OsbZIP46 activity in the presence of SAPK9 and OsPP2C6 dependent on ABA concentration and expression level. By using TGERP, we could characterize successfully the effects of ABA dependent gene expression signaling components in rice. In conclusion, TGERP represents very useful technology to study systemic functional genomics in rice or other monocots.

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Dynamic Behavior Analysis of an Eccentric Rotor with Unbalanced Magnetic Forces in BLDC Motors (BLDC 전동기의 전자기적 불평형력을 고려한 편심 회전자의 동적 거동 해석)

  • Kim, Tae-Jong;Hwang, Sang-Mun;Park, No-Gil
    • The Transactions of the Korean Institute of Electrical Engineers B
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    • v.48 no.11
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    • pp.604-610
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    • 1999
  • Vibration of a rotor-bearing system driven by an electric motor is a coupled phenomenon between mechanical characteristics and magnetic origins through the air-gap. With the advent of new high-energy magnets together with high precision motor applications, magnetic sources of vibration are becoming more serious. This paper investigates the transient whirl responses of a rotor system with purely mechanical origins and compares it with that of magnetically coupled origins. A perturbation method is applied to model the magnetic field associated with rotor eccentricity. Electromagnetic forces are obtained by the Maxwell stress method, which utilizes the analytical expression of radial flux density distribution. The FEM was applied to a rotor-motor system to illustrate magnetically coupled effects in rotor dynamics. Results show that magnetically coupled sources significantly affect the vibration of the rotor-motor system.

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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.

Transient and Steady State Analysis considering Roughness and Reynolds Number in Water Distribution Systems (관조도와 난류를 고려한 부정류와 정상류 해석의 적용 연구)

  • Kim, Hyunsu;Song, Youngsuk;Kim, Sanghyun
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.3
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    • pp.357-366
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    • 2006
  • In order to compute pressure variation for a water distribution system, an expression for the friction factor as a function of Reynolds number and the relative roughness needs to be properly incorporated in computational algorithm. Considering Moody s friction variation, Developed Unsteady Network Analyzer (UNA) has been modified to match computational results with EPANET 2.0. Substantial improvement can be found in the application of Improved UNA to both an hypothetical pipeline network and a real system located in Ulsan City. Random number generator is employed to represent the uncertainty of water use in real pipeline network. Comparisons of application between EPANET 2.0 and improved UNA 2.0 indicate advantages and potentials of this approach.

Experimental and Analytical Study on the Bus Duct System for the Prediction of Temperature Variations Due To the Fluctuation of Load

  • Thirumurugaveerakumar, S.;Sakthivel, M.;Valarmathi, S.
    • Journal of Electrical Engineering and Technology
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    • v.9 no.6
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    • pp.2036-2041
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    • 2014
  • In this paper, a thermal model is developed for the bus bar system to predict the temperature variation during the transient time period and to calculate both the steady-state and transient electrical current carrying capacity (ampacity) of bus bar. The bus bar system installed in the power house of Kumaraguru College of Technology, Coimbatore has been considered. Temperature variation predicted in the modelling is validated by observing the current and steady state temperatures in different feeders of the bus bar. Magnetic field of the extreme phases R and B induces more current in the middle phase Y. Hence, the steady state temperature in the phase Y is greater than other two phases. The transient capabilities of the bus bar are illustrated by calculating the variations in the bus bar temperature when it is subjected to a step change in current during the peak hours due to increase in hostel utilities and facilities (5.30 pm to 10.30 pm). The physical and geometrical properties of the bus bar and temperature variation in the bus bar are used to estimate the thermal time constants for common bus bar cross-sections. An analytical expression for the time constant of the bus bar is derived.

Introduction and Expression of Foreign Genes in Rice Cells by Particle Bombardment

  • Jeon, Jong-Seong;Jung, Hou-Sung;Sung, Soon-Kee;Lee, Jong-Seob;Choi, Yang-Do;Kim, Han-Jip;Lee, Kwang-Woong
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.27-36
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    • 1994
  • For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highest GUS gene expression of about 1000 blue spots per g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 110 mm target distance, 13 mm syringe filter holder and 5 $\mu$L DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron construct gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.

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The Effect of Cucumber mosaic virus 2b Protein to Transient Expression and Transgene Silencing Mediated by Agro-infiltration

  • Choi, Min-Sue;Yoon, In-Sun;Rhee, Yong;Choi, Seung-Kook;Lim, Sun-Hyung;Won, So-Youn;Lee, Yeon-Hee;Choi, Hong-Soo;Lee, Suk-Chan;Kim, Kook-Hyung;Lomonossoff, George;Sohn, Seong-Han
    • The Plant Pathology Journal
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    • v.24 no.3
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    • pp.296-304
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    • 2008
  • The transient and rapid expression system of a foreign protein in planta is a very useful technique in biotechnology application. We have investigated optimum condition of Agrobacterium-infiltration technique in which expression level of foreign proteins were maximized without detrimental effects on plants using GFP and Cucumber mosaic virus 2b protein, which is known as an enhancer of gene expression and a suppressor of post-transcriptional gene silencing(PTGS). The optimum expression level of both RNA and protein of GFP with minimum leaf impairment was obtained at $OD_{600}$=0.2 of Agrobactrium inocula. The steady-state levels of GFP RNA and protein generally peaked at 3 and 7 days post-infiltration(dpi), respectively. In the presence of 2b, both the magnitude and duration of GFP expression was highly increased and we could detect GFP level until 17 dpi. On the other hands, the 2b-mediated higher accumulation of foreign proteins resulted in the repression of normal leaf growth, possibly due to the limitation of supply of energy or materials required for growth maintenance. Using this Agrobacterium-infiltration system with 2b and GFP, we tested a hypothesis for the threshold model of PTGS initiation. Four GFP transgenic lines of N. benthamiana, which shows different expression level of GFP were tested to determine the threshold level for PTGS initiation. Agrobacterium-infiltration of GFP into those GFP-transgenic plants resulted in the co-silencing of the transgenic GFP. It was found that very low concentration of Agrobacterium with GFP and GFP+2b($OD_{600}$=0.002-0.02) which could not phenotypically induce an additive GFP expression, was enough to trigger PTGS pathway in all GFP transgenic plants. This strongly indicates that each GFP-transgenic plant should be expressing the transgenic GFP at its own pre-determined level and there was no buffer zone of additive GFP-expression to the threshold. In other words, the PTGS seems to be immediately activated as a self-defensive mechanism if an internal balance of gene expression is broken.

Uptake and Expression of Foreign Genes Using Seed-Derived Embryos of Rice (벼 종자 유래 배에서 외래유전자의 도입과 발현)

  • 정구흥
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.77-83
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    • 1994
  • DNA uptake in dry embryos of rice by DNA imbibition was detected by monitoring the expression of chimeric vectors. The selective markers of expression vectors used were ${\beta}-glucuronidase$ ronidase (GUS) and hygromycin phosphotransferase (HPT) genes under the control of CaMV35 S promoter. Frequency of transient expression of the foreign gene was generally 30-50% varying according to the types of vectors and rice cultivars. Dot blot analysis and DNA sequence analysis of inverse polymerase chain reaction products showed that selected rice in hygromycin B (HmB) medium had HPT gene and CaMV35S promoter DNA sequence in genomic DNA of rice. To investigate what ratio of rice having two marker genes simultaneously as rice embryos imbibed the vector DNA having two HPT and GUS gene, transform ants selected in lImB medium were subjected to PCR for GUS gene. It was shown that about 90 percentage of surviving ones in HmB medium had GUS gene.S gene.

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