• 약학회지
• /
• 제41권4호
• /
• pp.533-537
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제4권4호
• /
• pp.237-244
• /
• 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.

• 한국진공학회지
• /
• 제5권4호
• /
• pp.278-283
• /
• 1996

Field terminator를 이용하여 singl path cylindrical mirror analyser를 제작하였다. 먼저 컴퓨터 모의실험을 바탕으로 CMA의 원통형 전극양단에서 fringing field effect를 최소로 하여 log-scale의 등전위면을 그대로 유지하는 terminator의 전압비와 위치 등을 구하였다. 계산된 전압을 직렬로 연결된 금속산화물저항으로 만든 voltage divider로 field terminator에 직접 인가하여 CMA의 fringing field effect를 줄이는 방법으로 개발하였다. 이 방법으로 제작된 CMA의 분해능이 $\Delta$E/E=0.4% 이상임을 확인하였다.

• 대한전자공학회:학술대회논문집
• /
• 대한전자공학회 2002년도 ITC-CSCC -3
• /
• pp.1571-1574
• /
• 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.

• 한국해양공학회지
• /
• 제36권2호
• /
• pp.132-142
• /
• 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 d_x (x-directional distance between adjacent rows) within this WEC on its performance. In particular, this study enforces at least two equal d_x values within the 4×8 terminator array of vertical cylinders. It shows that a small value of this d_x leads to better energy extraction efficiency in some of these various d_x arrays than that of a corresponding regular array with the same d_x.

• BMB Reports
• /
• 제28권5호
• /
• pp.458-463
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제14권4호
• /
• pp.810-815
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제22권12호
• /
• pp.1714-1723
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• 제17권4호
• /
• pp.579-585
• /
• 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.

• Applied Biological Chemistry
• /
• 제44권4호
• /
• pp.211-214
• /
• 2001

Drosophila melanogaster Schneider line 2(S2)세포의 외래 단백질 발현 시스템을 이용한 외래 단백질의 한시적 발현을 검토하고, 서로 다른 terminator를 이용하였을 때의 단백질 발현 및 mRNA 발현 정도를 검토하였다. 한시적 발현의 경우 transfection agent를 제거하고 36-48시간 동안 배양한 경우, 가장 높은 green fluorescent protein(GEP)의 발현을 보였다. SV4O p(A), SV4O small T-antigen, 인간 gastrin 3'UTR을 terminator로 지니는 발현 벡터시스템에 각각 endostatin유전자를 cloning시킨 뒤 재조합 endostatin의 mRNA의 발현 정도를 비교하였다. 한시적 발현을 시킨 뒤 36시간 후 endostatin의 발현 정도를 비교해 본 결과 SV40 p(A)를 terminator로 사용했을 때 mRNA및 단백질의 발현이 가장 높았다.