• Title/Summary/Keyword: Tool-chain

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Insertion Mutation in HMG-CoA Lyase Increases the Production Yield of MPA through Agrobacterium tumefaciens-Mediated Transformation

  • Dong, Yuguo;Zhang, Jian;Xu, Rui;Lv, Xinxin;Wang, Lihua;Sun, Aiyou;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1924-1932
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    • 2016
  • Mycophenolic acid (MPA) is an antibiotic produced by Penicillium brevicompactum. MPA has antifungal, antineoplastic, and immunosuppressive functions, among others. ${\beta}-Hydroxy-{\beta}-methylglutaryl-CoA$ (HMG-CoA) lyase is a key enzyme in the bypass metabolic pathway. The inhibitory activity of HMG-CoA lyase increases the MPA biosynthetic flux by reducing the generation of by-products. In this study, we cloned the P. brevicompactum HMG-CoA lyase gene using the thermal asymmetric interlaced polymerase chain reaction and gene walking technology. Agrobacterium tumefaciens-mediated transformation (ATMT) was used to insert a mutated HMG-CoA lyase gene into P. brevicompactum. Successful insertion of the HMG-CoA lyase gene was confirmed by hygromycin screening, PCR, Southern blot analysis, and enzyme content assay. The maximum MPA production by transformants was 2.94 g/l. This was 71% higher than wild-type ATCC 16024. Our results demonstrate that ATMT may be an alternative practical genetic tool for directional transformation of P. brevicompactum.

The Study of Korean -type Discountstore Service Quality Scales(KD-SQS) (한국형 할인점의 서비스품질 측정 척도에 관한 연구)

  • Rho, Eun-Jeong;Suh, Yong-Gu
    • Journal of Distribution Research
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    • v.13 no.3
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    • pp.127-154
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    • 2008
  • For the large scale retailers such as chain discount stores and specialty stores, having measurement tool to maintain service quality can be very crucial to sustain high and same level of service qualities around multiple stores. This study aims to develop service quality scales of large-scale stores using Korean discount store cases. We have studied the previous service quality measures including RSQS(Retail Service Quality Scale) and have modified and developed our own scale model called KD-SQS(Korea Discount Service Quality Scale). The following six components have been revealed as the basic dimensions of service quality of Korean discount stores: basic benefits, promotion, personal interactions, physical aspects, policy and additional convenience. Our scales have been verified through various validity and reliability tests.

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A Study on Socio-Economic Effects of Italian Ports (이탈리아 항만의 사회경제학적 효과에 대한 연구)

  • D'agostini, Enrico;Ryoo, Dong-Keun
    • Journal of Navigation and Port Research
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    • v.41 no.4
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    • pp.243-250
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    • 2017
  • Seaports are not longer considered to be single entities for which the main activity is to load and discharge cargo, but rather as fundamental hubs within a complex supply chain serving global production networks. From a public perspective, they hold a key role in terms of their economic impact at the local, regional and national level by generating value added activity and employment. This issue is becoming increasingly pivotal for ports and their stakeholders. The objective of the study is to estimate a regression model of the value added activity and employment figures of 17 Italian ports for which there are no publications available. Concerning port system reform in Italy, the research intends to provide policy makers with a tool to measure the effects produced by ports and their importance to local communities and regions. The paper finds that in all Italian ports, the direct effects are larger than the indirect effects, and the bigger socio-economic effects are found in the biggest ports.

Application of Life Cycle Assessment to Enhance the Environmental Performance of Process Systems and Products (공정시스템과 제품의 환경성을 향상시키기 위한 전과정평가의 활용)

  • Lim, Seong-Rin
    • Clean Technology
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    • v.20 no.4
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    • pp.339-348
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    • 2014
  • Life cycle assessment (LCA) has become an important tool used to enhance the environmental performance of process systems and products. LCA is an essential element in design for environment (DfE) because LCA can be utilized to evaluate and analyze environmental impacts incurred in the life cycle and supply chain. This review presents methodologies that can be used to integrate LCA into DfE activities and reduce environmental impacts from process systems and products; and introduces case studies for water supply systems and cellular phones. LCA is first used to quantify environmental impacts and identify the principal contributor to high impacts. In the next step, environmental impacts from principal contributors can be reduced by using mathematical optimization tools as an engineering and technological approach and by utilizing the cooperation of professionals from a diverse range of fields. Because the methodologies and case studies can be applied and extended to other fields, this review paper can contribute to helping prevent environmental pollution and enhance the sustainability of our society.

Effect of cell-penetrating peptide-conjugated estrogen-related receptor ${\beta}$ on the development of mouse embryos cultured in vitro

  • Yang, Ning Jie;Seol, Dong-Won;Jo, Junghyun;Jang, Hyun Mee;Yoon, Sook-Young;Lee, Dong Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.1
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    • pp.1-8
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    • 2014
  • Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

Identification, Characterization, and DNA Sequencing of Nosema bombi in Bumblebees from Gangwon Province, Korea

  • Kwak, Kyu-Won;Yoon, Hyung-Joo;Choi, Youngcheol;Park, Kwanho;Hwang, Jaesam;Kim, Hyunae;Nam, Sunghee
    • International Journal of Industrial Entomology and Biomaterials
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    • v.27 no.2
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    • pp.219-227
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    • 2013
  • The purpose of this study was to quickly and effectively identify the Nosema disease of bumblebees in Gangwon Province in Korea. Bumblebees are crucial pollinators of various crops, and microsporidia are the critical pathogens of these hosts. When bumblebees are infected with Nosema bombi, their abdomens can become distended. Paralyzed and infected workers often become sluggish and die early. We have identified the morphology of the microsporidium by light and electron microscopy, and found it to have fairly small oval spores, as has been described previously in many other articles. For the specific and sensitive diagnosis of the microsporidian parasite N. bombi in bumblebees, we have developed an improved method of the polymerase chain reaction (PCR) for expeditious diagnosis. Two pairs of primers were tested on N. bombi and the related microsporidia Nosema apis and Nosema sp., both of which infect Bombus ignitus and Bombus hypocrita sapporoensis. Furthermore, we have verified and analyzed the 16SrRNA sequence data of N. bombi in bumblebees by using the Basic Local Alignment Search Tool (BLAST) server at the National Center for Biotechnology Information.

The Antifungal Test: An Efficient Screening Tool for the Discovery of Microbial Metabolites with Respiratory Inhibitory Activity

  • Han, Jae Woo;Kim, Bomin;Oh, Mira;Choi, Jaehyuk;Choi, Gyung Ja;Kim, Hun
    • Mycobiology
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    • v.48 no.4
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    • pp.326-329
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    • 2020
  • Valuable natural compounds produced by a variety of microorganisms can be used as lead molecules for development of new agrochemicals. Furthermore, high-throughput in vitro screening systems with specific modes of action can increase the probability of discovery of new fungicides. In the current study, a rapid assay tested with various microbes was developed to determine the degree of respiratory inhibition of Saccharomyces cerevisiae in two different liquid media, YG (containing a fermentable carbon source) and NFYG (containing a non-fermentable carbon source). Based on this system, we screened 100 fungal isolates that were classified into basidiomycetes, to find microbial secondary metabolites that act as respiratory inhibitors. Consequently, of the 100 fungal species tested, the culture broth of an IUM04881 isolate inhibited growth of S. cerevisiae in NFYG medium, but not in YG medium. The result is comparable to that from treatment with kresoxim-methyl used as a control, suggesting that the culture broth of IUM04881 isolate might contain active compounds showing the inhibition activity for respiratory chain. Based on the assay developed in this study and spectroscopic analysis, we isolated and identified an antifungal compound (-)-oudemansin A from culture broth of IUM04881 that is identified as Oudemansiella venosolamellata. This is the first report that (-)-oudemansin A is identified from O. venosolamellata in Korea. Taken together, the development of this assay will accelerate efforts to find and identify natural respiratory inhibitors from various microbes.

Determination of Short-term Bioconcentration Factor on Carbofuran in Carassius auratus (goldfish) (Carassius auratus(goldfish)를 이용한 Carbofuran의 단기간 생물농축계수의 측정)

  • 민경진;배영규;차춘근;박천만;강회양
    • Journal of Environmental Health Sciences
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    • v.22 no.4
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    • pp.25-32
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    • 1996
  • The Bioconcentration factor(BCF) is used as an important criterion in the risk assessment of environmental contaminants. Also it can be used as indicator of biomagnification of environmentally hazardous chemicals through food-chain as well as a tool for ranking the bioconcentration potential of the chemicals in the environment. This paper reports the measured BCF value on carbofuran in Carassius auratus(goldfish), under steady state, and examined corelation between the BCF value and the depuration rate constant. Carassius auratus(goldfish) was chosen as test organism and test periods were 1-day, 3-day and 5-day. Experimental concentrations were 0.05, 0.10 and 0.50 ppm. Carbofuran in fish tissue and in test water was extracted with n-hexane and acetonitril. GC-ECD was used to detect and quantitate carbofuran. The depuration rate of carbofuran from the whole body of goldfish is determined over the 24-h period after treatment. The obtained results were as follows: 1. It was possible to determine short term BCFs of carbofuran through relatively simple procedure in environmental concentrations. 2. $BCF_1$ of carbofuran in concentration of 0.05, 0.10 and 0.50 ppm were 1.66, 1.64 0.61, $BCF_3$ were 2.08, 2.14, 0.66 and $BCF_5$ were 2.21, 2.57, 0.86, respectively. 3. Carbofuran concentration in fish extract was increased as increasing test concentration and prolonging test period, but $BCF_s$ in concentration of 0.50 ppm was greately decreased. 4. Determined deputation rate constants of carbofuran in concentration of 0.05, 0.10, 0.50 ppm were 0.076, 0.082 and 0.089, respectively. 5. It is considered that great decrease of $BCF_s$ in concentration of 0.50 ppm is due to high water solubility and stability of carbofuran in testwater. 6. It is suggested that low BCF of carbofuran is due to its relatively high water solubility and depuration rate, compared to BPMC, carbaryl and chlorothalonil.

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Design and Implementation of a C-to-SystemC Synthesizer (C-to-SystemC 합성기의 설계 및 구현)

  • You, Myoung-Keun;Song, Gi-Yong
    • Journal of the Institute of Convergence Signal Processing
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    • v.10 no.2
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    • pp.141-145
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    • 2009
  • A C-to-SystemC synthesizer which processes the input behavior according to high-level synthesis, and then transforms the synthesis result into SystemC module code is implemented in this paper. In the synthesis process, the input behavioral description in C source code is scheduled using list scheduling algorithm and register allocation is performed using left-edge algorithm on the result of scheduling. In the SystemC process, the output from high-level synthesis process is transformed into SystemC module code by combining it with SystemC features such as channels and ports. The operation of the implemented C-to-SystemC synthesizer is validated through simulating the synthesis of elliptic wave filter in SystemC code. C-to-SystemC synthesizer can be used as a part of tool-chain which helps to implement SystemC design methodology covering from modeling to synthesis.

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Effect of Endocrine Disruptors on the Oocyte Maturation and Ovulation in Amphibians, Rana dybowskii

  • Choi, Mee-Jeong;Kim, Seung-Chang;Kim, An-Na;Kwon, Hyuk-Bang;Ahn, Ryun-Sup
    • Animal cells and systems
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    • v.11 no.1
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    • pp.1-8
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    • 2007
  • Recently, we have shown that some endocrine disruptors, heavy metals, organotins and azoles suppressed steroidogenic enzymes such as P450 side-chain cleavage enzyme (P450scc) and aromatase in bullfrog ovarian follicles. In the present study, by using an amphibian ovarian follicle culture system, we examined the effects of these endocrine disruptors on maturation and ovulation of oocytes from Rana dybowskii in vitro. Ovarian fragments or isolated follicles were cultured for 24 h in a medium containing frog pituitary homogenate (FPH) or progesterone ($P_{4}$) with or without endocrine disruptors, and oocyte maturation (germinal vesicle breakdown, GVBD) and ovulation were examined. Among the organotins, tributyltin (TBT) strongly inhibited both FPH-and $P_{4}-induced$ oocyte maturation ($ED_{50}$:0.6 and 0.7 ${\mu}M$, respectively); however, tetrabutyltin (TTBT) and dibutyltin (DBT) showed only partial suppression, while monobutyltin (MBT) showed no inhibitory effect. All of the organotins suppressed $P_{4}-induced$ oocyte ovulation very effectively at a low concentration, and TBT and DBT exerted an inhibitory effect on FPH-induced ovulation. Among the heavy metals, mercury (Hg), cadmium (Cd) and cobalt (Co) were very effective in inhibiting FPH-induced oocyte maturation and ovulation, while lead (Pb), arsenite (As) and zinc (Zn) were less effective. However, all of the heavy metals suppressed FPH-induced oocyte ovulation at a high dose ($100{\mu}M$). Among the azoles, itraconazole (ICZ), ketoconazole (KCZ) and clotrimazole (CTZ) effectively inhibited FPH-induced oocyte maturation and ovulation, while econazole (ECZ), miconazole (MCZ) and fluconazole (FCZ) were considerably less effective. These results demonstrated that the abovementioned endocrine disruptors exhibited differential effects on oocyte maturation and ovulation in amphibian follicles and that the frog ovarian culture system could be used as an effective experimental tool to screen and evaluate the toxicity of various endocrine disruptors in vitro.