• Title/Summary/Keyword: Selectable

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The Function of ArgE Gene in Transgenic Rice Plants

  • Guo, Jia;Seong, Eun-Soo;Cho, Joon-Hyeong;Wang, Myeong-Hyeon
    • Korean Journal of Plant Resources
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    • v.20 no.6
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    • pp.524-529
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    • 2007
  • We carried out to study the function of ArgE in transgenic rice plants, which were confirmed by PCR analysis and hygromycin selection. Transgenic rice plants were with selectable marker gene(HPT) inserted in genome of the rice. Southern analysis with hpt probe confirmed by two restriction enzymes that copy numbers of the selectable gene was introduced into the plant genome. We displayed that the relationship between drought stress and ArgE gene with the overexpressing rice plants. From this result, we observed that the degree of leaves damage has no difference in control and transgenic lines. The total RNAs were extracted from 6 weeks-seedling in normal condition in order to examine their expression levels with ArgE-overexpressed transgenic rice. In particular, expression patterns of genes encoding enzymes involved in abiotic stress, including drought and salt stresses. OsGF14a and OsSalt were investigated by reverse transcription-PCR(RT-PCR). Expression levels of the OsSalt gene decreased significantly in transgenic rice plants compared to control plant. However, ion leakage measurement did not demonstrate any leaves damage change between control and ArgE transgenic plants exposure to mannitol treatment. These results suggest that expression of the ArgE is not involved in tolerance for drought stress in rice but may playa role of signaling networks for salt-induced genes.

Experimental investigation of a new steel friction device with link element for seismic strengthening of structures

  • Papadopoulos, Panikos K.;Salonikios, Thomas N.;Dimitrakis, Stergios A.;Papadopoulos, Alkis P.
    • Structural Engineering and Mechanics
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    • v.46 no.4
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    • pp.487-504
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    • 2013
  • In the present work a new friction device, with a set of single or double rotational friction flanges and a link element, is described and tested. The mechanism may be applied for the strengthening of existing r/c or steel buildings as well as in new constructed buildings. The device has selectable variable behavior in different levels of displacement and an interlock mechanism that is provided by the link element. The link element may be designed to lock at preselected level of displacement, offering in this way an extra safety reserve against strong earthquakes. A summary of the existing literature about other similar mechanisms is initially presented in this paper. The proposed mechanism is presented and described in details. Laboratory experiments are presented in detail and the resulted response that proves the efficiency of the mechanism at selectable levels of strength capacity is discussed. Drawings of the mechanism attached to a r/c frame with connection details are also included. Finally a dynamic analysis of two r/c frames, with and without the proposed mechanism attached, is performed and the resulted response is given. The main conclusion is that the proposed mechanism is a cheap and efficient devise for the improvement of the performance of new or existing framed buildings to seismic loads.

One-Step Engineering of a Stable, Selectable Marker-Free Autoluminescent Acinetobacter baumannii for Rapid Continuous Assessment of Drug Activity

  • Jiang, Huofeng;Gao, Yamin;Zeng, Sheng;Wang, Shuai;Cao, Zhizhong;Tan, Yaoju;Yin, Huancai;Liu, Jianxiong;Zhang, Tianyu
    • Journal of Microbiology and Biotechnology
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    • v.29 no.9
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    • pp.1488-1493
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    • 2019
  • The rising cases of multidrug-resistant Acinetobacter baumannii (Ab) and the lack of effective drugs call for quick attention. Here, based on a Tn7 transposon and Xer/dif system, we constructed a stable, selectable marker-free autoluminescent Ab capable of producing visible light without extra substrates. Utilization of this autoluminescent reporter strain has the potential to reduce the time, effort and costs required for the evaluation of activities of anti-Ab drug candidates in vitro.

Efficient Gene Targeting using Nuclear Localization Signal (NLS) and Negative Selection Marker Gene in Porcine Somatic Cells

  • Kim, Hye Min;Lee, Sang Mi;Park, Hyo Young;Kang, Man-Jong
    • Reproductive and Developmental Biology
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    • v.38 no.2
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    • pp.71-77
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    • 2014
  • The specific genetic modification in porcine somatic cells by gene targeting has been very difficult because of low efficiency of homologous recombination. To improve gene targeting, we designed three kinds of knock-out vectors with ${\alpha}1,3$-galactosyltransferase gene (${\alpha}1,3$-GT gene), DT-A/pGT5'/neo/pGT3', DT-A/NLS/pGT5'/neo/pGT3' and pGT5'/neo/ pGT3'/NLS. The knock-out vectors consisted of a 4.8-kb fragment as the 5' recombination arm (pGT5') and a 1.9-kb fragment as the 3' recombination arm (pGT3'). We used the neomycin resistance gene (neo) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. These vectors have a neo gene insertion in exon 9 for inactivation of ${\alpha}1,3$-GT locus. DT-A/pGT5'/neo/pGT3' vector contain only positive-negative selection marker with conventional targeting vector. DT-A/NLS/pGT5'/neo/pGT3' vector contain positive-negative selection marker and NLS sequences in upstream of 5' recombination arm which enhances nuclear transport of foreign DNA into bovine somatic cells. pGT5'/neo/pGT3'/NLS vector contain only positive selection marker and NLS sequence in downstream of 3' recombination arm, not contain negative selectable marker. For transfection, linearzed vectors were introduced into porcine ear fibroblasts by electroporation. After 48 hours, the transfected cells were selected with $300{\mu}g/ml$ G418 during 12 day. The G418-resistant colonies were picked, of which 5 colonies were positive for ${\alpha}1,3$-GT gene disruption in 3' PCR and southern blot screening. Three knock-out somatic cells were obtained from DT-A/NLS/ pGT5'/neo/pGT3' knock-out vector. Thus, these data indicate that gene targeting vector using nuclear localization signal and negative selection marker improve targeting efficiency in porcine somatic cells.

MIC with Ripple Voltage Compensator Using Regenerative Snubber (에너지 회생형 스너브를 이용한 리플 전압 억제기를 가지는 MIC(Module Integrated Converter))

  • Kim, Hong-Sung;Chang, Hun Ki;Yoon, Yeo Young
    • Proceedings of the KIPE Conference
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    • 2014.11a
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    • pp.75-76
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    • 2014
  • This study deals with a isolation type MIC(Module Integrated Converter) for AC module with advantages such as DC Wireless, freely selectable installation capacity, minimized shadow effect etc. In this paper, MIC circuit with the function which can remove the ripple voltage of PV module and give the discharging path for charged energy in leakage inductance of isolation transformer. The validity of proposed circuir is verified by the simulation with PSIM.

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Construction of Plasmid Vectors for Zymomonas mobilis (Zymomonas mobilis의 Plasmid Vector 제조에 관한 연구)

  • Hwang, Duk-Ju;Rhee, Sang-Ki;Pack, Moo-Young
    • Microbiology and Biotechnology Letters
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    • v.15 no.5
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    • pp.319-327
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    • 1987
  • In order to develop useful plasmid vectors for Zymomonas cells, attempts were made to isolate natural plasmids from Z. mobilis ATCC10988. Among a few plasmids isolated, a small plasmid of 3.9 Kb size was chosen and designated as pZM3. By introducing the replication origin of pZM3 into pBR325, a hybrid plasmid vector of 8.4 Kb size, pHZ22, was constructed. This vector contained chloramphenicol resistant gene as a selectable marker and proved to be conjugally transmissible and stably maintained in Z. mobilis. Tetracycline resistant gene was isolated from RP4 and introduced into pHZ22 to make a new vector called pHZT224 of 10.7 Kb size. Through n series of experiments, it was evident that these plasmid vectors containing selectable markers of chloramphenicol and tetracycline resistance were shuttle vectors functional in Z. mobilis as well as E. coli.

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Transformation of Potato using the Phosphinothricin Acetyltransferase Gene as the Selectable Marker Gene (감자의 형질전환을 위한 표지유전자로서 Phosphinothricin Acetyltransferase 유전자의 이용)

  • Jeong, J.H.;Yang, D.C.;Bang, K.S.;Han, S.S.
    • Korean Journal of Weed Science
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    • v.18 no.3
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    • pp.205-213
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    • 1998
  • This experiment was carried out to produce herbicide resistant potatoes hawing only chimeric phosphinothricin acetyltransferase (PAT) genes without using antibiotic selectable marker. The pDY502 vector having only PAT gene was reconstructed for transformation of potato. The reconstructed vector was introduced to Agrobacterium tumefaciens MP90 disarmed, and they were used for potato transformation. Hormonal requirement for plant regeneration from leaves and stem explants of potato was investigated. From this experiment, MS medium treated with IBA 0.1 mg/L + BA 0.5 mg/L was the best for potato regeneration, and the ratio of shoot regeneration was 54% for leaf and 46% for stem in that condition. For transformation, explants of potato leaves and stems were cocultured with A. tumefaciens MP90 containing reconstructed vector harvoring only PAT gene. When the potato explants were placed on various concentrations of bialaphos and all the potato explants were dead on medium with over 5.0mg/L bialaphos. By this selection methods, the explants cocultured with Agrobacterium produced the putative transgenic shoots on medium with 5mg/L bialaphos treatment after 3-4 weeks. Second selection was performed by transferring the shoot tips of putative transgenic to medium containing 20mg/L of bialaphos. The shoot tips grew well on the second selection medium, indicating the production of successful transgenic plants. But normal shoots were dead in same cytotoxic medium. Incorporation of the PAT gene into transgenic potatos were confirmed by PCR analysis of DNA and Southern hybridization. These results show that the PAT gene can serve as a selectable marker and herbicide resistant genes for transformation of potato.

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Dosimetric Effect on Selectable Optimization Parameters of Volumatric Modulated Arc Therapy (선택적 최적화 변수(Selectable Optimization Parameters)에 따른 부피적조절회전방사선치료(VMAT)의 선량학적 영향)

  • Jung, Jae-Yong;Shin, Yong-Joo;Sohn, Seung-Chang;Kim, Yeon-Rae;Min, Jung-Wan;Suh, Tae-Suk
    • Progress in Medical Physics
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    • v.23 no.1
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    • pp.15-25
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    • 2012
  • The aim of this study is to evaluate plan quality and dose accuracy for Volumetric Modulated Arc Therapy (VMAT) on the TG-119 and is to investigate the effects on variation of the selectable optimization parameters of VMAT. VMAT treatment planning was implemented on a Varian iX linear accelerator with ARIA record and verify system (Varian Mecical System Palo Alto, CA) and Oncentra MasterPlan treatment planning system (Nucletron BV, Veenendaal, Netherlands). Plan quality and dosimetric accuracy were evaluated by effect of varying a number of arc, gantry spacing and delivery time for the test geometries provided in TG-119. Plan quality for the target and OAR was evaluated by the mean value and the standard deviation of the Dose Volume Histograms (DVHs). The ionization chamber and $Delta^{4PT}$ bi-planar diode array were used for the dose evaluation. For treatment planning evaluation, all structure sets closed to the goals in the case of single arc, except for the C-shape (hard), and all structure sets achieved the goals in the case of dual arc, except for C-shape (hard). For the variation of a number of arc, the simple structure such as a prostate did not have the difference between single arc and dual arc, whereas the complex structure such as a head and neck showed a superior result in the case of dual arc. The dose distribution with gantry spacing of $4^{\circ}$ was shown better plan quality than the gantry spacing of $6^{\circ}$, but was similar results compared with gantry spacing of $2^{\circ}$. For the verification of dose accuracy with single arc and dual arc, the mean value of a relative error between measured and calculated value were within 3% and 4% for point dose and confidence limit values, respectively. For the verification on dose accuracy with the gantry intervals of $2^{\circ}$, $4^{\circ}$ and $6^{\circ}$, the mean values of relative error were within 3% and 5% for point dose and confidence limit values, respectively. In the verification of dose distribution with $Delta^{4PT}$ bi-planar diode array, gamma passing rate was $98.72{\pm}1.52%$ and $98.3{\pm}1.5%$ for single arc and dual arc, respectively. The confidence limit values were within 4%. The smaller the gantry spacing, the more accuracy results were shown. In this study, we performed the VMAT QA based on TG-119 procedure, and demonstrated that all structure sets were satisfied with acceptance criteria. And also, the results for the selective optimization variables informed the importance of selection for the suitable variables according to the clinical cases.