• Title/Summary/Keyword: terminator

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Characteristics of the Resistance Phenotypes by Inducible Resistance Gene ermK and Its Terminator Region Mutants (MLS계 항생제에 대한 유도내성 유전자 ermK 및 그 돌연변이체의 유도내성 표현형)

  • Choi, Sung-Sook;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.533-537
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    • 1997
  • The characteristics of the resistance phenotypes of Bacillus subtilis having ermk and its terminator region mutants were determined. Wild type ermK(pEC101) and pECMT109(methylase SD-region mutant) showed typical inducible resistance phenotype. pECMF1(terminator1 region mutant) and pECMT2(terminator2 region mutant) showed constitutive resistance to Kitasamycin but inducible resistance to tylosin. In contrast, pECMT3(terminator1 and terminator2 double mutant) and pECMT309(terminator1, terminator2 and methylase SD region triple mutant) showed constitutive resistance both to kitasamycin and tylosin.

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Effect of Transcription Terminators on Expression of Human Lipocortin-1 in Recombinant Saccharomyces cerevisiae

  • Chung, Bong-Hyun;Kim, Byung-Moon;Nam, Soo-Wan;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.237-244
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    • 1994
  • The vector systems for the expression and secretion of human lipocortin-l (LC1) from Saccharomyces cerevisiae were constructed with GAL10 promoter and the prepro leader sequence of mating factor-$\alpha$1. They were further constructed to contain three different transcription terminators; GAL7 terminator, LCl terminator and a fused form of these two terminators. The expression and secretion levels of LCl were compared to investigate the effect of transcription terminators on the LCl gene expression. For the expression cassettes employing the GAL7 terminator or the terminator of fused form, the expression levels of LCl were measured by scanning the immunoreactive LCl protein bands, and were found to be 0.27 g/l and 0.32 g/l, respectively. The highest expression level of 0.54 g/l was obtained with the expression vector containing the LCl transcription terminator. In all expression cassettes, the majority of LCl proteins expressed were retained intracellularly, indicating a low secretion efficiency of about 5%. The high expression level of LCl was explained by the great content and stability of LCl mRNA transcribed from the LCl terminator-employing vector. The results of this study demonstrate that the LCl transcription terminator functions for the expression of LCl in S. cerevisiae better than the GAL7 terminator.

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The Study on CMA using field terminator (Field terminator를 이용한 CMA 제작에 관한 연구)

  • 이충만;성면창;권순남;정광호
    • Journal of the Korean Vacuum Society
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    • v.5 no.4
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    • pp.278-283
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    • 1996
  • We constructed a single path cylindrical mirror analyser(CMA) using field terminator methods. With computer simulation, the best fit voltage ratio and position of the field terminaor was determined with maintain log-scale equipotential line near both end of the CMA. Then we construct field terminator with voltage divider of metal-oxide resisters which reduces the fringe field effect. The resolving power of the CMA was better than $\Delta$E/E=0.4%.

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Programmable Digital On-Chip Terminator

  • Kim, Su-Chul;Kim, Nam-Seog;Kim, Tae-Hyung;Cho, Uk-Rae;Byun, Hyun-Guen;Kim, Suki
    • Proceedings of the IEEK Conference
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    • 2002.07c
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    • pp.1571-1574
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    • 2002
  • This paper describes a circuit and its operations of a programmable digital on-chip terminator designed with CMOS circuits which are used in high speed I/O interface. The on-chip terminator matches external reference resistor with the accuracy of ${\pm}$ 4.1% over process, voltage and temperature variation. The digital impedance codes are generated in programmable impedance controller (PIC), and the codes are sent to terminator transistor arrays at input pads serially to reduce the number of signal lines. The transistor array is thermometer-coded to reduce impedance glitches during code update and it is segmented to two different blocks of thermometer-coded transistor arrays to reduce the number of transistors. The terminator impedance is periodically updated during hold time to minimize inter-symbol interferences.

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Numerical Investigation of Multi-body Wave Energy Converters' Configuration

  • Heo, Kyeonguk;Choi, Yoon-Rak
    • Journal of Ocean Engineering and Technology
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    • v.36 no.2
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    • pp.132-142
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    • 2022
  • We investigate the performance of multi-body wave energy converters (WECs). This investigation considers multiple scattering of water waves by the buoys of a WEC under the generalized mode approach. Predominantly, the effect of a WEC's configuration on its energy extraction is studied in this research. First, single-row terminator and single-column attenuator arrays of vertical cylinders have been studied. The performance of these attenuator arrays shows that the wall effect induced by the periodic buoys influences the wave propagation and energy extraction in these WECs. Further studies show that a single-row terminator array of vertical cylinders performs better than the corresponding single-column attenuator array. Subsequently, multi-row terminator arrays of vertical cylinders are investigated by conducting a parametric study. This parametric study shows that the hydrodynamic property of three resonance phenomena makes energy extraction efficiency drop down, and the magnitude of energy extracted oscillates between the resonance points in these WECs. Finally, a 4×8 terminator array of vertical cylinders is studied to determine the effect of various dx (x-directional distance between adjacent rows) within this WEC on its performance. In particular, this study enforces at least two equal dx values within the 4×8 terminator array of vertical cylinders. It shows that a small value of this dx leads to better energy extraction efficiency in some of these various dx arrays than that of a corresponding regular array with the same dx.

Disruptions of Two Apparent rho-Independent Transcription Terminator Structures do not help in Enhancing the Expression of aceK in E. coli

  • Lee, Su-Ji;Chung, Taeo-Wan
    • BMB Reports
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    • v.28 no.5
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    • pp.458-463
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    • 1995
  • Two apparent rho-independent transcription terminator structures within the coding sequence of aceK have been destroyed to access their roles in the differential expression between aceA and aceK in the glyoxylate bypass operon of E. coli. The effect of mutations on the expression of aceK was evaluated in two different ways: one by maxicell labeling and the other by lacZ fusion gene construction. The maxicell labeling experiment with the mutant operon clones has failed, like that of the wild type operon clone, to visibly show isocitrate dehrogenase (IDH) kinase/phosphatase, the product of aceK, on the autoradiogram of a protein gel. When the same mutations were introduced into an aceK::lacZ fusion gene to quantitatively evaluate the mutational effect, the activity of ${\beta}-galactosidase$ in neither of the mutant versions of the fusion gene was elevated significantly enough to explain the degree of polarity observed in this region. Thus, we conclude that neither of these intragenic, apparent rho-independent transcription terminator structures, which have long been suspected as a major determinant in the down regulation of aceK, really act as a premature transcriptional terminator.

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High-Level Expression of Recombinant Human Interleukin-2 in Chinese Hamster Ovary Cells Using the Expression System Containing Transcription Terminator

  • Kim, Eun-Ju;Kim, Dong-Jun;Hwang, Hye-Yeon;Yoon, Jae-Seung;Yoon, Ye-Up;Baek, Kwang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.810-815
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    • 2004
  • Many biological properties and the clinical potential of human interleukin-2 (hIL-2) draw much attention to its high-level expression in mammalian cells. Recombinant human IL-2 (rhIL-2) was expressed in Chinese hamster ovary (CHO) cells, using the recently developed expression system which confers position-independent expression. Stable CHO cell lines carrying several hundred amplified copies of the rhIL-2 gene were easily obtained and rhIL-2 was expressed at high levels after selection with increasing concentrations of methotrexate. Interestingly, the insertion of the transcription terminator of the human gastrin gene into the downstream region of the gene for rhIL-2 considerably increased rhIL-2 expression. Using the expression system with the transcription terminator, it was possible to get a CHO cell line expressing the rhIL-2 at a very high level, about $11.4\mug/10^6$ cell/day, which is about 6 times higher than that previously reported. The biological activity of the rhIL-2 protein purified from the cell line was also confirmed by the cell proliferation assay.

Identification of the ${\beta}$-Glucosidase Gene from Bifidobacterium animalis subsp. lactis and Its Expression in B. bifidum BGN4

  • Youn, So Youn;Park, Myeong Soo;Ji, Geun Eog
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1714-1723
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    • 2012
  • ${\beta}$-Glucosidase is necessary for the bioconversion of glycosidic phytochemicals in food. Two Bifidobacterium strains (Bifidobacterium animalis subsp. lactis SH5 and B. animalis subsp. lactis RD68) with relatively high ${\beta}$-glucosidase activities were selected among 46 lactic acid bacteria. A ${\beta}$-glucosidase gene (bbg572) from B. lactis was shotgun cloned, fully sequenced, and analyzed for its transcription start site, structural gene, and deduced transcriptional terminator. The structural gene of bbg572 was 1,383 bp. Based on amino sequence similarities, bbg572 was assigned to family 1 of the glycosyl hydrolases. To overexpress bbg572 in Bifidobacterium, several bifidobacteria expression vectors were constructed by combining several promoters and a terminator sequence from different bifidobacteria. The maximum activity of recombinant Bbg572 was achieved when it was expressed under its own promoter and terminator. Its enzyme activity increased 31-fold compared with those of its parental strains. The optimal pH for Bbg572 was pH 6.0. Bbg572 was stable at $37-40^{\circ}C$. It hydrolyzed isoflavones, quercetins, and disaccharides with various ${\beta}$-glucoside linkages. Bbg572 also converted the ginsenosides Rb1 and Rb2. These results suggest that this new ${\beta}$-glucosidase-positive Bifidobacterium transformant can be utilized for the production of specific aglycone products.

Expression of Recombinant Human Growth Hormone in a Soluble Form in Escherichia coli by Slowing Down the Protein Synthesis Rate

  • Koo, Tai-Young;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.579-585
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    • 2007
  • Formation of inclusion bodies is usually observed when foreign proteins are overexpressed in E. coli. The formation of inclusion bodies might be prevented by lowering the rate of protein synthesis, and appropriate regulation of the protein expression rate may lead to the soluble expression. In this study, human growth hormone (rhGH) was expressed in a soluble form by slowing down the protein synthesis rate, which was controlled in the transcriptional and translational levels. The transcriptional level was controlled by the regulation of the amount of RNA polymerase specific to the promoter in front of the rhGH gene. For lowering the rate of translation, the T7 transcription terminator-deleted vector was used to synthesize the longer mRNA of the target gene because the longer mRNA is expected to reduce the availability of tree ribosomes. In both methods, the percentage of soluble expression increased when the expression rate slowed down, and more than 93% of rhGH expressed was a soluble form in the T7 transcription terminator-deleted expression system.

Effect of Transcriptional Terminator Sequences on Recombinant Protein Expression from Drosophila melanogaster S2 Cell (전사 종결 염기 서열이 Drosophila melanogaster Schneider line 2 세포에서 외래 단백질의 발현에 미치는 영향)

  • Hwang, In-Sook;Park, Jong-Hwa;Lee, Youn-Hyung;Yoon, Jae-Seung;Baek, Kwang-Hee;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.44 no.4
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    • pp.211-214
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    • 2001
  • Utilizing the foreign gene expression system of Drosophila melanogaster Schneider line 2(S2) cell, the degree of transient protein and mRNA expression was examined with different terminators. In the case of transient expression, the expression level of green fluorescent protein(GFP) was the highest when the transfection agent was eliminated and then cultivated for 36 to 48 hr. The terminators(SV40 p(A), SV40 small T-antigen and human gastrin 3'UTR) of the expression vector system were each cloned with endostatin; thereafter, the expression levels of the endostatin and its mRNA were compared. When the expression levels of endostatin were compared 36 hr after transfection, the SV40 p(A) terminator showed the highest expression level of endostatin and its mRNA.

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