• Title/Summary/Keyword: complementation

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Conditions for Intergeneric Protoplast Fusion of Yeast (효모의 이속간 원형질체 융합조건)

  • Kim, Young-Ho;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.383-389
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    • 1985
  • Optimum conditions of PEG treatment for the intergeneric fusion of yeast protoplasts were investigated. Fusants were selected by nutritional complementation on minimal medium. The intergeneric fusion frequency between pro-toplasts of S. cerevisiae and C. tropicalis was distributed 10$^{-4}$ to 10$^{-6}$, depending on the combination of parental strains. PEG 4000 or 6000 are equally effective. 30%(w/v) PEG 4000 was found to be optimum and below 20% its stabilizing effect was lost, resulting in protoplast lysis, and optimum pH was 8.0. The efficiency of PEG was enhanced by higher temperature of the PEG solution, and by the addition of Ca ions. The stimulating effect of Ca ions in the range of 1 mM to 100 mM proved similar.

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Transformation and Expression of the PAT Gene in Arabidopsis Tryptophan Mutants

  • Lim, Seon-Hee;Kim, Young-Soon;Cheong, Hyeon-Sook
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.243-247
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    • 1996
  • Phosphoribosylanthranilate transferase (PAT) catalyzes the second step of the tryptophan biosynthetic pathway and is encoded by a single-copy gene that complements all the visible phenotypes of the tryptophan mutant (trp1-100) of Arabidopsis. The trp1-100 is blue fluorescent under UV light becuase it accumulates anthranilate. To obtain a plant with reduced PAT activity, PAT1 genes with several internal deletions in different promoter regions (pHS 101, pHS102, pHS104, pHS105, and pHS107) were induced into trp1-100 via Agrobacterium. Then, homozygous T3 plants were isolated and examined for blue fluorescence. Introduction of the PAT1 gene fusants results in the reversion of fluorescence phenotype except in the case of pHS105. These results prompted us to perform a parallel analysis of anthranilate synthase and PAT interms of the genetic complementation. A plant line carrying pHS105 gene fusant does not completely complement the blue fluorescence but it accumulates less anthranilate than trp1-100. The activity of PAT was reduced in the transgenic mutant as well. The plant carrying these constructs will add to the growing collection of molecular tools for the study of the indolic secondary metabolism.

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Characterization of Virulence Function of Pseudomonas cichorii Avirulence Protein E1 (AvrE1) during Host Plant Infection

  • Huong, Duyen Do Tran;Rajalingam, Nagendran;Lee, Yong Hoon
    • The Plant Pathology Journal
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    • v.37 no.5
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    • pp.494-501
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    • 2021
  • Pseudomonas cichorii secretes effectors that suppress defense mechanisms in host plants. However, the function of these effectors, including avirulence protein E1 (AvrE1), in the pathogenicity of P. cichorii, remains unexplored. In this study, to investigate the function of avrE1 in P. cichorii JBC1 (PcJBC1), we created an avrE1-deficient mutant (JBC1ΔavrE1) using CRISPR/Cas9. The disease severity caused by JBC1ΔavrE1 in tomato plants significantly decreased by reducing water soaking during early infection stage, as evidenced by the electrolyte leakage in infected leaves. The disease symptoms caused by JBC1ΔavrE1 in the cabbage midrib were light-brown spots compared to the dark-colored ones caused by PcJBC1, which indicates the role of AvrE1 in cell lysis. The avrE1-deficient mutant failed to elicit cell death in non-host tobacco plants. Disease severity and cell death caused by JBC1ΔavrE1 in host and non-host plants were restored through heterologous complementation with avrE1 from Pseudomonas syringae pv. tomato DC3000 (PstDC3000). Overall, our results indicate that avrE1 contributes to cell death during early infection, which consequently increases disease development in host plants. The roles of PcJBC1 AvrE1 in host cells remain to be elucidated.

For/from Alternations in Causative 'FOR/FROM V-ing' Constructions ('For/From V-ing' 사역구문의 전치사 for/from 교체현상 연구)

  • Kim, Mija
    • Cross-Cultural Studies
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    • v.49
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    • pp.1-32
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    • 2017
  • This paper discusses the structural and meaning features of causative from/for V-ing constructions as complement and provides insight on their grammatical characteristics revealed from alternation between prepositions for and from in nonfinite V-ing complement clause constructions. Guided by empirical data, this paper demonstrates that there are three types of syntactic patterns classified by the main verbs in these constructions and that these three syntactic types are closely linked with the meaning. These classifications are supported by the passivizations and aspect. In addition, this paper suggests that the function of for and from followed by nonfinite V-ing clause should be treated as a preposition introducing nonfinite V-ing clauses.

Genome-wide Screening to Identify Responsive Regulators Involved in the Virulence of Xanthomonas oryzae pv. oryzae

  • Han, Sang-Wook;Lee, Mi-Ae;Yoo, Youngchul;Cho, Man-Ho;Lee, Sang-Won
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.84-89
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    • 2019
  • Two-component systems (TCSs) are critical to the pathogenesis of Xanthomonas oryzae pv. oryzae (Xoo). We mutated 55 of 62 genes annotated as responsive regulators (RRs) of TCSs in the genome of Xoo strain PXO99A and identified 9 genes involved in Xoo virulence. Four (rpfG, hrpG, stoS, and detR) of the 9 genes were previously reported as key regulators of Xoo virulence and the other 5 have not been characterized. Lesion lengths on rice leaves inoculated with the mutants were shorter than those of the wild type and were significantly restored with gene complementation. The population density of the 5 mutants in planta was smaller than that of PXO99A at 14 days after inoculation, but the growth curves of the mutants in rich medium were similar to those of the wild type. These newly reported RR genes will facilitate studies on the function of TCSs and of the integrated regulation of TCSs for Xoo pathogenesis.

Curcumin modulates the apolipoprotein B mRNA editing by coordinating the expression of cytidine deamination to uridine editosome components in primary mouse hepatocytes

  • He, Pan;Tian, Nan
    • The Korean Journal of Physiology and Pharmacology
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    • v.23 no.3
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    • pp.181-189
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    • 2019
  • Curcumin, an active ingredient of Curcuma longa L., can reduce the concentration of low-density lipoproteins in plasma, in different ways. We had first reported that curcumin exhibits hypocholesterolemic properties by improving the apolipoprotein B (apoB) mRNA editing in primary rat hepatocytes. However, the role of curcumin in the regulation of apoB mRNA editing is not clear. Thus, we investigated the effect of curcumin on the expression of multiple editing components of apoB mRNA cytidine deamination to uridine (C-to-U) editosome. Our results demonstrated that treatment with $50{\mu}M$ curcumin markedly increased the amount of edited apoB mRNA in primary mouse hepatocytes from 5.13%-8.05% to 27.63%-35.61%, and significantly elevated the levels of the core components apoB editing catalytic polypeptide-1 (APOBEC-1), apobec-1 complementation factor (ACF), and RNA-binding-motif-protein-47 (RBM47), as well as suppressed the level of the inhibitory component glycine-arginine-tyrosine-rich RNA binding protein. Moreover, the increased apoB RNA editing by $50{\mu}M$ curcumin was significantly reduced by siRNA-mediated APOBEC-1, ACF, and RBM47 knockdown. These findings suggest that curcumin modulates apoB mRNA editing by coordinating the multiple editing components of the edito-some in primary hepatocytes. Our data provided evidence for curcumin to be used therapeutically to prevent atherosclerosis.

Characteristics of civil anti-disaster organization in Korean Disaster Management Systems : focusing on CAIND (citizen corps active in disaster) (국내 민간 방재조직의 특성에 관한 고찰 -지역자율방재단을 중심으로-)

  • Kim, Bo-Kyun;Chae, Jong-Sik
    • The Korean Journal of Emergency Medical Services
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    • v.25 no.1
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    • pp.135-146
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    • 2021
  • Purpose: This study aimed to evaluate the Citizen-Corps-Active-In-Disaster (CAIND) according to a new viewpoint based on changes in basic theoretical concepts and establish a development plan. Methods: A documentary survey was administered, and the study followed the technical approach method. Detailed information on CAIND was taken from scholarly monographs, specialty publications, and previous studies, including master and doctorate dissertations. Results: CAIND may be defined as legal auxiliary organizations composed of local residents who volunteer to prepare for "all kinds of disasters as well as prevention activities" and instill the ideology of "leading service organizations of safety as a form of community involvement," "resident-friendly organizational activities based on autonomous operation," and "performing activities which supplement their demands according to the realization of social welfare and regional conditions." Conclusion: The newly established basic theoretical concept included the concepts of "professionalism and possibilities," "social expandability," "voluntary participation," and "the realization and complementation of social welfare"

A Study on the Application of Private Security in Assembly and Demonstration Sites Using SNS Emotional Language Analysis

  • Kunhee, Kim;Cheolyeung, Jang
    • International Journal of Advanced Culture Technology
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    • v.10 no.4
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    • pp.1-7
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    • 2022
  • The purpose of this study is to review the appropriateness of deploying private security guards using SMS emotional language analysis at the current assembly and demonstration sites, which rely only on the police force, and to suggest a plan. Therefore, it is intended to contribute to suppressing the problems at the assembly and demonstration sites and present a new paradigm for responding to the assembly and demonstration sites based on the study's results. First, it is necessary to prepare a legal basis for the deployment of private security guards in the 'Security Services Industry Act' and 'Assembly and a Demonstration Act'. Second, there is a need for a 'security company selection criteria' for selecting security companies with a lot of experience, such as collective civil petition sites and security for national critical facilities (special security services). Third, it is necessary to prepare financial resources for the deployment of private security guards. This study is expected to serve as a new cornerstone for effective management of assembly and demonstration sites through mutual complementation of the police and private security.

Expression and Activity of Citrus Phytoene Synthase and $\beta$-Carotene Hydroxylase in Escherichia coli

  • Kim, In-Jung;Ko, Kyong-Cheol;Nam, Tae-Sik;Kim, Yu-Wang;Chung, Won-Il;Kim, Chan-Shick
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.212-218
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    • 2003
  • Citrus phytoene synthase (CitPsy) and ${\beta}$-carotene hydroxylase (CitChx), which are involved in caroteinoid biosynthesis, are distantly related to the corresponding bacterial enzymes from the point of view of amino acid sequence similarity. We investigated these enzyme activities using Pantoea ananatis carotenoid biosynthetic genes and Escherichia coli as a host cell. The genes were cloned into two vector systems controlled by the T7 promoter. SDS-polyacrylamide gel electrophoresis showed that CitPsy and CitChx proteins are normally expressed in E. coli in both soluble and insoluble forms. In vivo complementation using the Pantoea ananatis enzymes and HPLC analysis showed that ${\beta}$-carotene and zeaxanthin were produced in recombinant E. coli, which indicated that the citrus enzymes were functionally expressed in E. coli and assembled into a functional multi-enzyme complex with Pantoea ananatis enzymes. These observed activities well matched the results of other researchers on tomato phytoene synthase and Arabidopsis and pepper ${\beta}$-carotene hydroxylases. Thus, our results suggest that plant carotenoid biosynthetic enzymes can generally complement the bacterial enzymes and could be a means of carotenoid production by molecular breeding and fermentation in bacterial and plant systems.