• Title/Summary/Keyword: Cell Transformation

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The Effects of Co-cultivation Medium and Culture Conditions on Rice Transformation Efficiency (공동배양과정의 배지조성과 배양조건이 벼 형질전환효율에 미치는 영향)

  • Kim, Yul-Ho;Park, Hyang-Mi;Choi, Man-Soo;Yun, Hong-Tai;Choi, Im-Soo;Shin, Dong-Bum;Kim, Chung-Kon;Lee, Jang-Yong
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.252-260
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    • 2009
  • Rice is the most important cereal crop not only in supplying the basic staple food for more than half of the world's population but also as a model plant for functional genomic studies of monocotyledons. Although rice transformation method using A. tumefaciens has already been widely used to generate transgenic plants, the transformation rate is still low in most Korean elite cultivars. We made several modifications of the standard protocol especially in the co-cultivation step to improve the efficiency of the rice transformation. The co-culture medium was modified by the addition of three antioxidant compounds (10.5 mg/L L-cysteine, 1 mM sodium thiosulfate, 1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (5 mg/L silver nitrate). Co-cultivation temperature ($23.5^{\circ}C$ for 1 day, $26.5^{\circ}C$ for 6 days) and duration (7 days) were also changed. The plasmid of pMJC-GB-GUS carrying the GUS reporter gene and the bar gene as the selectable marker was used to evaluate the efficiency of the transformation. After co-cultivation, a high level of GUS gene expression was observed in calli treated with the modified method. It is likely that those newly added compounds helped to minimize the damage due to oxidative bursts during plant cell-Agrobacterium interaction and to prevent necrosis of rice cells. And the transformation rate under the modified method was also remarkably increased approximately 8-fold in Heungnambyeo and 2-fold in Ilmibyeo as compared to the corresponding standard method. Furthermore, we could produce the transgenic plants stably from Ilpumbyeo which is a high-quality rice but its transformation rate is extremely low. Transformation and the copy number of transgenes were confirmed by PCR, bar strip and Southern blot analysis. The improved method would attribute reducing the effort and the time required to produce a large number of transgenic rice plants.

Signaling for Synergistic Activation of Natural Killer Cells

  • Kwon, Hyung-Joon;Kim, Hun Sik
    • IMMUNE NETWORK
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    • v.12 no.6
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    • pp.240-246
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    • 2012
  • Natural killer (NK) cells play a pivotal role in early surveillance against virus infection and cellular transformation, and are also implicated in the control of inflammatory response through their effector functions of direct lysis of target cells and cytokine secretion. NK cell activation toward target cell is determined by the net balance of signals transmitted from diverse activating and inhibitory receptors. A distinct feature of NK cell activation is that stimulation of resting NK cells with single activating receptor on its own cannot mount natural cytotoxicity. Instead, specific pairs of co-activation receptors are required to unleash NK cell activation via synergy- dependent mechanism. Because each co-activation receptor uses distinct signaling modules, NK cell synergy relies on the integration of such disparate signals. This explains why the study of the mechanism underlying NK cell synergy is important and necessary. Recent studies revealed that NK cell synergy depends on the integration of complementary signals converged at a critical checkpoint element but not on simple amplification of the individual signaling to overcome intrinsic activation threshold. This review focuses on the signaling events during NK cells activation and recent advances in the study of NK cell synergy.

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.

Optimized Cell ID Codes for SSDT Power Control in W-CDMA System (W-CDMA 시스템의 최적의 SSDT 전력 제어용 셀 식별 부호)

  • Young-Joon Song;Bong-Hoe Kim;Hae Chung
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.13 no.8
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    • pp.804-810
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    • 2002
  • The code division multiple access(CDMA) system capacity is limited by the amount of interference of the system. To reduce the unnecessary interference, this paper proposes optimized cell identification codes for site selection diversity transmission(SSDT) power control in wideband code division multiple access system of third generation partnership project(3GPP). The main objective of SSDT power control is to transmit on the downlink from the primary cell, and thus reducing the interference caused by the multiple transmission. In order to select a primary cell, each cell is assigned a temporary identification(ID) and user equipment(UE) periodically informs a primary cell ID to the connecting cells during soft handover. The non-primary cells selected by UE do not transmit the dedicated physical data channel(DPDCH) to reduce the interference. A major issue with the SSDT technology is the impact of uplink symbol errors on its performance. These errors can corrupt the primary ID code and this may lead to wrong decoding in the base station receivers. The proposed SSDT cell ID codes are designed to minimize the problem and to be easily decoded using simple fast Hadamard transformation(FHT) decoder.

Translation Initiation Factor 4E (eIF4E) is Regulated by Cell Death Inhibitor, Diap1

  • Lee, Sun Kyung;Lee, Ji Sun;Shin, Ki Soon;Yoo, Soon Ji
    • Molecules and Cells
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    • v.24 no.3
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    • pp.445-451
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    • 2007
  • Translation initiation factor 4E (eIF4E) is a key regulator of protein synthesis. Abnormal regulation of eIF4E is closely linked to oncogenic transformation. Several regulatory mechanisms affecting eIF4E are discussed, including transcriptional regulation, phosphorylation and binding of an inhibitor protein. However it is not clear how the level of eIF4E protein is regulated under basal conditions. Here we demonstrate that Diap1 (Drosophila Inhibitor of Apoptosis Protein), a cell death inhibitor, binds directly to eIF4E and poly-ubiquitinates it via its E3 ligase activity, promoting its proteasome-dependent degradation. Expression of Diap1 caused a reduction of Cyclin D1 protein level and inhibited the growth stimulation induced by overexpression of eIF4E. Taken together, our results suggest that the level of eIF4E protein is regulated by Diap1, and that IAPs may play a role in cap-dependent translation by regulating the level of eIF4E protein.

Studies on the Requirements of $Ca^{++}$ for Cell Division and $Ca^{++}$ Permeability of Plasma Membrane of Fast Dividing Mouse Embryo Cells. (세포분열이 왕성한 생쥐 배세포에서 세포분열에 대한 $Ca^{++}$의 요구와 세포막투과성에 대한 연구)

  • Bae, In-Ha;Park, Ji-Hye
    • Clinical and Experimental Reproductive Medicine
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    • v.14 no.2
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    • pp.93-100
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    • 1987
  • To determine the effect of calcium on the preimplantational development of mouse two-cell embryo, the various concentrations of calcium were added into the culture media and the rate of blastocyst formation was observed. Also, to examine the effect of trifluoperazine, an inhibitor of calmodulin which is involved in the several intracellular calcium functions, embryos were cultured for 48 hours at the various concentrations of this inhibitor. An additional 24 hour culture was done to examine the effect of this drug on the transformation from morula to blastocyst. The results are as following ; 1. About 1.71mM of extracellular calcium is adequate for blastocyst formation and the higher concentrations of calcium (3.43mM and 8.55mM) do not affect on the blastocyst formation and the degenerating rate. 2. Trifluoperazine $100{\mu}M$ presents the inhibitory effect on the blastocyst formation while $1{\mu}M$ and $10{\mu}M$ do not so. 3. After an additional 24 hour culture, there is transformation of morula to blastocyst and the degenerating rate of embryo is increased all together.

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Comparison of the Cell Surface Barrier and Enzymatic Modification System in Brevibacterium flavum and B. Lactofermentum

  • Jang Ki-Hyo;Britz Margaret L.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.225-229
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    • 2005
  • To investigate impediments to plasmid transformation in Brevibacterium flavum BF4 and B. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties. B. lactofermentum was more sensitive to growth inhibition by glycine than B. flavum. Release of cellular proteins during sonication was more rapid for B. lactofermentum than for B. flavum. Plasmid DNA (pCSL 17) isolated from B. flavum transformed recipient $McrBC^+$ strains of Escherichia coli with lower efficiency than $McrBC^-$. McrBC digestion of this DNA confirmed that B. flavum contain methylated cytidines in the target sequence of McrBc sequences but B. lactofermentum contained a different methylation pattern. DNA derived from the B. lactofermentum transformed recipient $EcoKR^+$ strains of E. coli with lower efficiency than $EcoKR^-$, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.

Removal of Aerosol Through Fibrous Filter as a Function of Particle Size and Velocity (입자의 크기와 유속에 따른 섬유질 여과포에 의한 부유입자 제거 연구)

  • Sang Hoon Lee;Kwan Sik Chun;Hoo Keun Lee
    • Nuclear Engineering and Technology
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    • v.16 no.1
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    • pp.29-35
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    • 1984
  • Filtration efficiency tests were conducted on a fiberglass mat filter with DOP aerosol having a diameter from 0.l${\mu}{\textrm}{m}$ to 0.45${\mu}{\textrm}{m}$ in the face velocity range of 1cm/sec to 10cm/sec. Filtration of submicron particles by a fibrous filter is characterized by a face velocity. The size of DOP aerosol which has a minimum removal efficiency decreases with increasing the velocity. A numerical solution of the diffusion equation is obtained for a fiberglass mat filter by using "Kuwabara's cell model" for the flow field and Von Mises Transformation for the actual flow around a fiberglass. The present theoretical results agree quite well with the experimentals for fiberglass mat. This result could be contributed to predict the removal efficiency on an air filter and to optimize the operating condition of an air purification system with a filter. a filter.

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Studies on the Induction of Transformation in Cereal Plants. III. Cultures and Regeneration of Rice Protoplasts Transferred Foreign Genes. (곡물류의 형질전환 유도에 관한 연구 III. 외래 유전자가 도입된 벼 원형질체의 배양 및 재분화)

  • Hwang, Baik;Hwang, Sung-Jin;Im, Hyong-Tak;Kang, Young-Hee
    • KSBB Journal
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    • v.8 no.1
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    • pp.62-68
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    • 1993
  • Transformed rice plantlet were recovered from protoplasts by electroporation with the plasmld pB 1121, which contain the plant expressible NPT-II and GUS genes. Embryonic cell suspension culture was established with embryonic callus induced from mature seeds of rice (Oryza sativa L. cv. Dong-jin) on the MS medium supplemented with 2.0 mg/l 2,4-D, 0.5 mg/l kinetin, 3% sucrose. Protoplasts isolated from embryonic cell suspensions were electroplated and then poterltialty-transformed tissues were selected by growth on the medium containing 200 mg/l kanamycin sulfate. When subjected to GUS assay, they stained blue, indicating the expression of the inserted GUS genes. Plantlets were regenerated from electroplated protoplasts on the hormone free MS medium. Transferred foreign genes in the plants were confirmed by southern hybridization. These results support use of electroporation for transformation of these important cereal plants.

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Studies on Cellular Factors Responsible for 2,3,7,8-TCDD Resistency and Cellular Transformation (2,3,7,8-TCDD의 세포형질전환 및 내성획득에 관여하는 세포내 인자에 관한 연구)

  • Ryeom Tai-Kyung;Choi Young-Sill;Kim Ok-Hee;Kang Ho-Il
    • Environmental Mutagens and Carcinogens
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    • v.26 no.1
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    • pp.1-6
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    • 2006
  • To enhance our understanding of toxicity mediated through the pathway by which TCDD stimulates gene expression, we have investigated genes whose expressions are changed after treatment with TCDD and/or MNNG in human Chang liver cell. First, we treated with MNNG and TCDD for two weeks to transform human Chang liver cell. We obtained cell looks like to be transformed and compared the differential gene expression by using cDNA chip (Macrogen) which carrys genes related with signal transduction pathways, oncogenes and tumor suppressor genes, etc. We found that TCDD up- or down-regulated 203 and 111 genes including oncogenes and tumor suppressor genes in human Chang liver cell two fold or more, respectively. Second, we compared the differential gene expression after treatment with TCDD only by using cDNA chip (Superarray) which carrys genes related with cell cycle regulations, and found that TCDD up regulated genes related with cell proliferation as well as cell growth inhibition in human Chang liver cell two fold or more, respectively. These results suggest that toxicity induced by TCDD may reflect sustained alterations in the expression of many genes and that the changes reflect both direct and indirect effects of TCDD.

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