• Journal of fish pathology
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• v.24 no.3
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• pp.297-305
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• 2011

A trap identification system for isolating functional sequences to allow the constitutive expression of foreign protein from Edwardsiella tarda was developed. Using the green fluorescent protein (GFP) reporter-based trap system, various functional sequences to drive heterologous expression of the GFP were selectable in Escherichia coli host. However from the bioinformatic sequence analysis, all the segments predicted as regulatory regions were not native promoters actually existing upstream of endogenous E. tarda genes. Instead, a number of non-authentic sequences, possibly resulted from the random shuffling and/or intermolecular ligation were also proven to be able to display a potent GFP expression in the recombinant E. coli. Further analysis with selected clones showed that both authentic and non-authentic sequences could function in as a constitutive promoter, leading quite a consistent and stable GFP expression after repetitive subcultures. Microscopic examination also confirmed the uniform pattern of GFP expression in every host bacterium. Semi-quantitative assay of GFP showed that there was no clear relationship between expression levels and organizational features of the promoters trapped. Functional promoter-like elements achieved in the present study could be a good starting material for multivalent genetic engineering of E. tarda in order to produce recombinant vaccines in a cost-effective fashion.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.105-109
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• 2005

Heat Shock Transcription Factor (HSF), and the promoter heat Shock Element (HSE), are among the most highly conserved transcriptional regulatory elements in nature. HSF mediates the transcriptional response of eukaryotic cells to heat, infection and inflammation, pharmacological agents, and other stresses. While HSF is essential for cell viability in yeast, oogenesis and early development in Drosophila, extended life-span in C. elegans, and extra-embryonic development and stress resistance in mammals, little is known about its full range of biological target genes. We used whole genome analyses to identify virtually all of the direct transcriptional targets of yeast HSF, representing nearly three percent of the genomic loci. The majority of the identified loci are heat-inducibly bound by yeast HSF, and the target genes encode proteins that have a broad range of biological functions including protein folding and degradation, energy generation, protein secretion, maintenance of cell integrity, small molecule transport, cell signaling, and transcription. Approximately 30% of the HSF direct target genes are also induced by the diauxic shift, in which glucose levels begin to be depleted. We demonstrate that phosphorylation of HSF by Snf1 kinase is responsible for expression of a subset of HSF targets upon glucose starvation.

• BMB Reports
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• v.41 no.3
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• pp.204-209
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• 2008

The cDNAs encoding glutathione peroxidase (GPx) were cloned and sequenced from the liver of three Chinese carps with different tolerance to hepatotoxic microcystins, phyto-planktivorous silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis), and herbivorous grass carp (Ctenopharyngodon idellus). Using genome walker method, a 750 bp 5'-flanking region of the silver carp GPx gene was obtained, and several potential regulatory elements were identified in the promoter region of the GPx gene. The silver carp GPx gene was widely expressed in all tissues examined. Despite phylogenetic analysis, assigning this newly described carp GPx to the group of mammalian GPx2, the carp GPx seems more similar to GPx1 from a physiological point of view. The constitutive expression pattern of the three carp liver GPx gene, shows a positive relationship with their tolerance to microcystins.

• BMB Reports
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• v.32 no.5
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• pp.456-460
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• 1999

The promoter of the HSV-1 VP16 gene contains binding sites for the cellular transcription factors such as USF, CTF, and Sp1, each of which affects basal level expression of the VP16 gene. Transcription of the VP16 gene was induced by viral immediate-early proteins, ICP0 and ICP4, in a synergistic manner but repressed by ICP22. To gain further insight into the role of ICP0 in the expression of the VP16 gene during virus infection, several mutants with deletions in each of their transcriptional regulatory elements were generated. According to transient gene expression assays of these mutants using the CAT gene as a reporter, the USF and CTF binding sites were necessary for efficient induction of the promoter in the presence of transfected ICP0 or during virus infection, whereas the Sp1 binding site had little effect on ICP0-mediated VP16 expression. These results indicate that the immediate early proteins of HSV-1 regulate expression of the VP16 gene during virus infection by modulating the activities of cellular transcription factors such as USF and CTF.

• Korean Journal of Animal Reproduction
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• v.18 no.2
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• pp.133-139
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• 1994

Two human lactoferrin expression vectors(pCChcLf and pCChcLf-1) were constructed using rat $\beta$-casein gene and human lactoferrin cDNA. The recombinant DNAs containing human lactoferrin cDNA were microinjected into the fertilized eggs of hybrid mice (BDF1 : C57BL$\times$DBA) and the DNA-injected eggs were treansferred into the oviducts of foster mothers. Genomic DNAs were isolated from the tails of mice born from the microinjected eggs and analyzed by Southern blot analysis. As a result, 5 and 9 transgenic mice with CChcLf and CChcLf-1 gene were produced, respectively. To determine tissue-specificity of transgene expression, Northern blot analysis was performed. Female transgenic mice were killed at day 10 of lactation and total RNAs from various tissues were isolated. Based on Northern blot analysis, it was shown that transgene was mainly expressed in the mammary glands of transgenic mice. In addition, the human lactoferrin in milk was detected by enzyme-linked immunosorbent assay. For this study, milk was obtained from the mammary glands of the transgenic mice at day 10 of lactation. In line #2 of CChcLf and line #7 of CChcLf-1 transgenic mice, human lactoferrin was secreted into the milk at concentration levels of 340ng/ml and 60ng/ml, respectively.

• Nuclear Engineering and Technology
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• v.52 no.7
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• pp.1471-1480
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• 2020

This research presents a case study on the remediation of a radioactive waste (uranium: U) utilizing a multi-objective fuzzy optimization in an electrocoagulation process for the iron-stainless steel and aluminum-stainless steel anode/cathode systems. The incorporation of the cumulative uncertainty of result, operational cost and energy consumption are essential key elements in determining the feasibility of the developed model equations in satisfying specific maximum contaminant level (MCL) required by stringent environmental regulations worldwide. Pareto-optimal solutions showed that the iron system (0 ㎍/L U: 492 USD/g-U) outperformed the aluminum system (96 ㎍/L U: 747 USD/g-U) in terms of the retained uranium concentration and energy consumption. Thus, the iron system was further carried out in a multi-objective analysis due to its feasibility in satisfying various uranium standard regulatory limits. Based on the 30 ㎍/L MCL, the decision-making process via fuzzy logic showed an overall satisfaction of 6.1% at a treatment time and current density of 101.6 min and 59.9 mA/㎠, respectively. The fuzzy optimal solution reveals the following: uranium concentration - 5 ㎍/L, cumulative uncertainty - 25 ㎍/L, energy consumption - 461.7 kWh/g-U and operational cost based on electricity cost in the United States - 60.0 USD/g-U, South Korea - 55.4 USD/g-U and Finland - 78.5 USD/g-U.

• Molecules and Cells
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• v.39 no.8
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• pp.581-586
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• 2016

Post-translational modifications (PTMs) of proteins are essential to increase the functional diversity of the proteome. By adding chemical groups to proteins, or degrading entire proteins by phosphorylation, glycosylation, ubiquitination, neddylation, acetylation, lipidation, and proteolysis, the complexity of the proteome increases, and this then influences most biological processes. Although small RNAs are crucial regulatory elements for gene expression in most eukaryotes, PTMs of small RNA microprocessor and RNA silencing components have not been extensively investigated in plants. To date, several studies have shown that the proteolytic regulation of AGOs is important for host-pathogen interactions. DRB4 is regulated by the ubiquitin-proteasome system, and the degradation of HYL1 is modulated by a de-etiolation repressor, COP1, and an unknown cytoplasmic protease. Here, we discuss current findings on the PTMs of microprocessor and RNA silencing components in plants.

• STI Policy Review
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• v.1 no.3
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• pp.1-15
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• 2010

This paper makes the case that a new policy strategy to enhance a global green recovery is needed urgently. The new strategy requires two essential elements. First, G20 economies should follow the lead of South Korea and China and turn their green stimulus investments into a serious long-term commitment, and to support these investments, they should adopt environmental pricing policies and instigate pricing and regulatory reforms to reduce carbon dependency. Second, the G20 also needs to target and coordinate assistance to developing economies in science, technology and innovation (STI) for clean energy. Such assistance is essential to help developing economies to overcome the skills, technological and capital gap that they face in clean energy technologies over the long term. Reform of the Clean Development Mechanism (CDM) is also necessary to establish a long-term global price signal for carbon, and to increase the coverage of developing economies, the sectors and technologies and the overall financing of clean energy projects. Formulating such a policy strategy should appeal to both the Asian-Pacific and Western economies comprising the G20, and by working together to formulate such a strategy, the G20 could lead the way toward a new era of global economic management and STI cooperation in clean energy.

• Journal of Plant Biotechnology
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• v.40 no.1
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• pp.27-36
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• 2013

We isolated and functionally characterized a Dendrobium Actin1 (DmACT1) promoter that drives strong gene expression in the orchid genus Dendrobium. A genomic fragment containing the region 3227 bp upstream of the coding region of DmACT1 was obtained by inverse PCR. Detailed comparison of the full-length cDNA and genomic sequences revealed that DmACT1 has a 1374 bp first intron in the 5' UTR. However, the 5' flanking sequences upstream of the coding region showed no obvious sequence similarities compared to those of known promoters, including plant actin promoters. Serial deletion constructs of the 5' flanking region from the translation initiation codon were fused to the coding sequence of a GUS/luciferase fusion reporter to identify the regulatory elements necessary for promoter activity. Transient assays in the flowers of Dendrobium revealed that the 5' UTR-intron greatly enhanced promoter activity. Moreover, the DmACT1 promoter with its 5' UTR-intron yielded approximately 10-fold higher reporter activity than the rice Act1 promoter-intron. Our data suggest that the DmACT1 promoter with its 5' UTR-intron is a useful tool for strong expression of transgenes in Dendrobium orchids.

• Journal of Radiation Protection and Research
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• v.28 no.4
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• pp.281-290
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• 2003

The KN-12 spent nuclear fuel transport cask, which is a Type B(U) package designed to comply with the requirements of Korea Atomic Energy Act[1], IAEA Safety Standards Series No.TS-R-1[2] and US 10 CFR Part 71[3], is designed for carrying up to 12 PWR spent fuel assemblies in a basket structure. The cask has been licensed in accordance with Korea Atomic Energy Act and was fabricated in Korea in accordance with the requirements of ASME B&PV Sec.III, Div.3[4]. The cask must maintain thermal integrity in accordance with the related regulations and be evaluated to verify that the thermal performance of the cask complies with the regulatory requirements. The temperatures of the cask and components were determined by using finite elements methods with a numerical tool, safety tests using an 1/8 height slice model of the real cask were conducted to demonstrate verification of the numerical tool and methods, and heat transfer tests for normal transport conditions were performed as a fabrication acceptance test to demonstrate the heat transfer capability of the cask.