• Title/Summary/Keyword: That-Complementation

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Characterization of RAD3 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD3 유사유전자의 분리와 특성)

  • Choi In Soon
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.1023-1027
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    • 2004
  • The RAD3 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. An yeast RAD3 gene has been previously isolated by functional complementation. In order to identify the RAD3 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD3 DNA, and then isolated RAD3 homologous DNA from C. cinereus chromosome. The RAD3 homolog DNA was contained in 3.2 kb DNA fragment. Here, we report the results of characterization of a fungus C. cinereus homolog to the yeast RAD3 gene. Southern blot analysis confirmed that the C. cinereus chromosome contains the RAD3 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from the C. cinereus cells were hybridized with the 3.4 kb PvuII DNA fragment of the S. cerevisiae RAD3 gene, transcripts size of 2.8 kb were detected. In order to investigate whether the increase of the amount of transcripts by DNA damaging agent, transcript levels were examined after treating agents to the cells. The level of transcripts were not increased by untraviolet light (UV). This result indicated that the RAD3 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the HRD3 gene is essential for viability of the cells and DNA repair function. These observations suggest an evolutionary conservation of other protein components with which HRD3 interacts in mediating its DNA repair and viability functions.

Pro-Apoptotic Role of the Human YPEL5 Gene Identified by Functional Complementation of a Yeast moh1Δ Mutation

  • Lee, Ji Young;Jun, Do Youn;Park, Ju Eun;Kwon, Gi Hyun;Kim, Jong-Sik;Kim, Young Ho
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.633-643
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    • 2017
  • To examine the pro-apoptotic role of the human ortholog (YPEL5) of the Drosophila Yippee protein, the cell viability of Saccharomyces cerevisiae mutant strain with deleted MOH1, the yeast ortholog, was compared with that of the wild-type (WT)-MOH1 strain after exposure to different apoptogenic stimulants, including UV irradiation, methyl methanesulfonate (MMS), camptothecin (CPT), heat shock, and hyperosmotic shock. The $moh1{\Delta}$ mutant exhibited enhanced cell viability compared with the WT-MOH1 strain when treated with lethal UV irradiation, 1.8 mM MMS, $100{\mu}M$ CPT, heat shock at $50^{\circ}C$, or 1.2 M KCl. At the same time, the level of Moh1 protein was commonly up-regulated in the WT-MOH1 strain as was that of Ynk1 protein, which is known as a marker for DNA damage. Although the enhanced UV resistance of the $moh1{\Delta}$ mutant largely disappeared following transformation with the yeast MOH1 gene or one of the human YPEL1-YPEL5 genes, the transformant bearing pYES2-YPEL5 was more sensitive to lethal UV irradiation and its UV sensitivity was similar to that of the WT-MOH1 strain. Under these conditions, the UV irradiation-induced apoptotic events, such as FITC-Annexin V stainability, mitochondrial membrane potential (${\Delta}{\psi}m$) loss, and metacaspase activation, occurred to a much lesser extent in the $moh1{\Delta}$ mutant compared with the WT-MOH1 strain and the mutant strain bearing pYES2-MOH1 or pYES2-YPEL5. These results demonstrate the functional conservation between yeast Moh1 and human YPEL5, and their involvement in mitochondria-dependent apoptosis induced by DNA damage.

Research on the Leadership Types in Italian Restaurants (이태리 레스토랑 종사자들의 리더십 유형에 관한 연구)

  • Yim, Seoung-Bean;Kim, Pan-Jin
    • Journal of Distribution Science
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    • v.10 no.12
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    • pp.35-43
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    • 2012
  • Purpose - This study analyzes the effects of types of leadership on the employees of Italian restaurants, its efficacy, and organizational citizenship behavior, utilizing a causal assessment model. In this study, independent variables such as the type of leadership perceived in the manager or chef by an Italian restaurant's employees, and its efficacy were parameters, and the organizational citizenship behavior and organizational effectiveness were the variables representing the results in the hypothesis. The study aimed to draw implications by verifying the leadership via efficacy and the impact on organizational citizenship behavior of Italian restaurants. Research design, data, methodology - For the purpose of this analysis, specific questionnaire items were configured according to the theory and efficacy of the study. From a questionnaire used in organizational citizenship behavior comprising 22 questions, six were modified to suit the research purpose of this study. The configured questionnaire comprised 5 parts and 40 items. A Likert (Likert) 5-point scale was utilized to measure responses to the questionnaire items from the employees of an Italian restaurant in Seoul who participated in the survey. For data collection, 400 questionnaires were distributed, and 344 collected. Factor analysis and reliability verification were conducted using SPSS18.0 and AMOS18.0. A covariance structure analysis was conducted to test the research hypotheses. Results - Based on the results of the analyses, the summary and suggested implications of the research are as follows: The covariance structure analysis used to analyze the kind of effect transformational and transactional leadership styles in Italian restaurant employees had on self-efficacy, group-efficacy, and organizational citizenship behavior, indicated that among the characteristics of transformational leadership (such as, idealized influence, inspirational motivation, individual consideration, and intellectual stimulation), idealized influence and individual consideration had a positive influence on self-efficacy. Idealized influence, individual consideration, conditional reward, and management by exception also positively influenced self-efficacy and altruistic and conscientious behavior (organizational citizenship behavior). Conclusions - Results suggest that with regard to self-efficacy and group efficacy, managers in different departments and chefs should provide team members with a vision for the future, increase their confidence in their abilities, and build their trust in the organization. By evaluating employee performance and experiences, management can demonstrate leadership and encourage organizational citizenship behavior through enjoyable, voluntary participation. Transformational and transactional leadership is effective in group processes that include social-exchange relationships, self-efficacy and group efficacy, and organizational citizenship behavior. However, as this research study utilizes only self-reported data, it has several limitations, such as a vulnerability of errors caused by the various experiment types. A significant limitation of this study is the lack of potential for the duplication of results. The covariance structure analysis, however, provides complementation to limit the impact of errors from self-reporting studies. A future study can extend this research by utilizing different data collection methods.

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Identification and Functional Analysis of the Chain Length Determinant Gene ste8 Involved in the Biosynthesis of Ebosin by Streptomyces sp. 139

  • Yang, Zhang;Li, Xiaohua;Qi, Xiaoqaing;Shan, Junjie;Jiang, Rong;Guo, Lianhong;Zhang, Ren;Li, Yuan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1500-1508
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    • 2013
  • Ebosin, a novel exopolysaccharide produced by Streptomyces sp. 139, has obvious antirheumatic arthritis activity in vivo, and its biosynthesis gene cluster (ste), consisting of 27 open reading frames, has been identified. This paper reports our study of the gene functionality of ste8, the predicted protein product of which is homologous to some bacterial chain length determinant Wzz proteins. For characterization of Ste8, ste8 was cloned and expressed in the mutant strain E. coli 086:H2 (${\Delta}wzz$). The functional complementation of wzz by ste8 was demonstrated by the restoration of wild-type lipopolysaccharide biosynthesis and increased levels of serum resistance of E. coli 086:H2 (${\Delta}wzz$) (pET30a-ste8). To examine the function of ste8 in ebosin biosynthesis, the gene was knocked out with a double crossover via homologous recombination. The molecular weight of the ebosin derivative EPS-8m produced by the mutant Streptomyces sp. 139 ($ste8^-$) was much lower than that of ebosin, and the binding activity of EPS-8m for IL-1R decreased significantly compared with ebosin. These results demonstrate that ste8 encodes a chain length determinant (Wzz) that functions in ebosin biosynthesis.

Molecular cloning and characterization of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (CaHDR) from Camptotheca acuminata and its functional identification in Escherichia coli

  • Wang, Qian;Pi, Yan;Hou, Rong;Jiang, Keji;Huang, Zhuoshi;Hsieh, Ming-shiun;Sun, Xiaofen;Tang, Kexuan
    • BMB Reports
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    • v.41 no.2
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    • pp.112-118
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    • 2008
  • Camptothecin is an anti-cancer monoterpene indole alkaloid. The gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (designated as CaHDR), the last catalytic enzyme of the MEP pathway for terpenoid biosynthesis, was isolated from camptothecin-producing Camptotheca acuminata. The full-length cDNA of CaHDR was 1686 bp encoding 459 amino acids. Comparison of the cDNA and genomic DNA of CaHDR revealed that there was no intron in genomic CaHDR. Southern blot analysis indicated that CaHDR belonged to a low-copy gene family. RT-PCR analysis revealed that CaHDR expressed constitutively in all tested plant organs with the highest expression level in flowers, and the expression of CaHDR could be induced by 100 ${\mu}M$ methyl-jasmonate (MeJA), but not by 100 mg/L salicylic acid (SA) in the callus of C. acuminata. The complementation of CaHDR in Escherichia coli ispH mutant MG1655 demonstrated its function.

Structural Analysis of Multi-Functional Fishway in Seomoon Weir (서문보의 다기능 어도의 구조해석)

  • Lee, Young Jae;Lee, Jung Shin;Jang, Hyung Kyu
    • Ecology and Resilient Infrastructure
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    • v.7 no.4
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    • pp.308-319
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    • 2020
  • In this study, the field applicability of the recently constructed multifunctional fishway in Seomunbo, Yeongcheon-si, and Gyeongsangbuk-do were examined. The analysis variables were R/C slab (S1) and R/C+S/C slab (S2), the underground passage standard areas (width × length) were 1.4 m × 0.2 m, 1.4 m × 0.3 m, and 1.4 m × 0.6 m, and the flow velocities were 0.8, 1.2, and 1.6 m/s. As a result of the analysis, the safety of the design of Seomunbo was evaluated. The analysis showed compared to the Seomoon Weir fishway, the maximum stress of S2 decreased by 24 - 32%, the bending moment of the underground passage decreased by 16 - 33%, the maximum stress of the sidewall decreased by 20 - 36%. In addition, the bending moment of the upper slab decreased by 17 - 33%, the maximum stress of the upper slab decreased by 9 - 28%, and the bending moment decreased by 19 - 33%. Complementation was required in the following percentages: 18% and 14% for the maximum stress and bending moment of the underground passage, respectively, 15% and 17% for the maximum sidewall stress and bending moment, respectively, and 11% and 16% for the upper slab maximum stress and bending moment, respectively. The results showed that S2 was superior to that of the Seomoon Weir fishway, and the underground passage size of 1.4 m × 0.3 m was superior to those of 1.4 m × 0.2 m and 1.4 m × 0.6 m, and R/C+S/C slab was superior to that of R/C slab. The findings are expected to be useful for constructing and designing the multifunctional fishway.

A Study on Policy Acceptance Intention to Use Artificial Intelligence-Based Public Services: Focusing on the Influence of Individual Perception & Digital Literacy Level (인공지능 기반 공공서비스 정책수용 의도에 관한 연구: 개인의 인식과 디지털 리터러시 수준이 미치는 영향을 중심으로)

  • Jang, Changki;Sung, WookJoon
    • Informatization Policy
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    • v.29 no.1
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    • pp.60-83
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    • 2022
  • The purpose of this study is to empirically analyze the effect of individual perception of artificial intelligence and the level of digital literacy on the acceptance of artificial intelligence-based public services. For empirical analysis, a research model was set up based on the technology acceptance model and planned behavior theory using survey data of 2017 and analyzed through structural equations. To summarize the results of the analysis, firstly, the positive perception of individuals about artificial intelligence technology plays a role in reinforcing attitudes toward benefits and reducing concerns about public service in which artificial intelligence technology has been introduced. Secondly, the level of digital literacy reinforces both benefits and concerns about artificial intelligence technology, but it was found that the intention to use public services was reinforced through the benefits of artificial intelligence technology perceived by individuals, rather than privacy concerns about artificial intelligence technology. Thirdly, it was confirmed that the perceived benefits of individuals on artificial intelligence technology reinforced the intention to use public civil services, and privacy concerns negatively influenced the intention to use. It was confirmed that the influence of a perceived ease of use and usefulness, as opposed to privacy concerns, further reinforces the intention to use. Both citizens' positive perceptions regarding the accuracy and reliability of information provided through artificial intelligence technology and institutional complementation of responsibility for errors caused by artificial intelligence technology are strengthened, and technical problems related to privacy protection are solved.

The Mouse Mutations Circling and Spinner are Allelic

  • Kyoung in Cho;Lee, Eun-Ju;Kim, Myoung-Ok;Kim, Sung-Hyun;Pakr, Jun-Hong;Jung, Boo-Kyung;Kim, Hee-Chul;Sol ha Hwang;Suh, Jun-Gyo
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.90-90
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    • 2003
  • Circling mice were recorded to display profound deafness and a head-tossing and bidirectional circling behavior, showing an autosomal recessive mode of inheritance. In addition, the histological examination of inner ears revealed that the region around organ of Corti, spiral ganglion neurons and outer hair cells showed definite abnormality. On the other hand, a genetic linkage map was constructed in an intraspecific backcross between cir and C57BL/6J mice. The cir gene was mapped to a region between D9Mitl16/D9Mit15 and D9Mit38 on the mouse chromosome 9. Estimated distances between cir and D9Mitl16, and between cir and D9Mit38 are 0.70 $\pm$ 0.40 and 0.23 $\pm$ 0.23 cM, respectively. The markers in order was defined as follows: centromere-D9Mit182- D9Mit51/ D9Mit79/ D9Mit310- D9Mit212/ D9Mit184- D9Mit116/ D9Mit15- cir- D9Mit38- D9Mit20- D9Mit243- D9Mit16- D9Mit55/ D9Mit125- D9Mit281 Based on genetic mapping, we constructed for a YAC contig across cir region. They covered the entire region or cir and cir gene was located on between the lactotransferrin (ltf) and the macrotubule-associated protein (map4). It is known that sr gene is localized in 64cM of mouse chromosome 9. The two mouse were found to be allelic by complementation test. Recently the spinner mouse has been mapped to our cir region, and tmie gene were elucidated. And further study will be needed in circling mouse to prove tmie gene mutaiton.

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Molecular Cloning and Nucleotide Sequence Analysis of pyrB Gene Encoding Aspartate Transcarbamylase from Psychrophilic Sporosarcina psychrophilia (저온성균 Sporosarcina psychrophilia로부터 Aspartate Transcarbamylase 유전자의 클로닝 및 염기서열 분석)

  • 성혜리;안원근;김사열
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.312-319
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    • 2002
  • The Sporosarcina psychrophilia pyrB gene, which encodes aspartate transcarbamylase (ATcase), was cloned on Sau3AI restriction endonuclease fragment inserted into pUC19 plasmid vector, S. psychrophilia pyrB gene was expressed in Escherichia coli pyrB mutant for the complementation test. The sequence of 2,606 nucleotides including putative pyrB gene was determined. The region contained one full open reading frame (ORf) and two partial ORFs. The deduced amino acid sequence of the second ORF showed 59% identity with that of Bacillus caldolyticus ATCase. The first and third partial ORFs were closely related to the uracil permease (pyrP) and dihydroorotase (pyrC), respectively. Besides, potential terminator, antiterminator, and anti-antiterminator structures were found in the intergenic region between pyrP and pyrB. These results suggested that S. psychrophilia pyrimidine nucleotide biosynthesis genes are clustered as well as other Bacillus sp. Over-expressed product of pyrB encoding ATCase was purified and analyzed by the SDS-PAGE. The purified PyrB protein turned out to be molecular mass of 27 kDa and showed ATCase activity.

The Line n-sigraph of a Symmetric n-sigraph-V

  • Reddy, P. Siva Kota;Nagaraja, K.M.;Geetha, M.C.
    • Kyungpook Mathematical Journal
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    • v.54 no.1
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    • pp.95-101
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    • 2014
  • An n-tuple ($a_1,a_2,{\ldots},a_n$) is symmetric, if $a_k$ = $a_{n-k+1}$, $1{\leq}k{\leq}n$. Let $H_n$ = {$(a_1,a_2,{\ldots},a_n)$ ; $a_k$ ${\in}$ {+,-}, $a_k$ = $a_{n-k+1}$, $1{\leq}k{\leq}n$} be the set of all symmetric n-tuples. A symmetric n-sigraph (symmetric n-marked graph) is an ordered pair $S_n$ = (G,${\sigma}$) ($S_n$ = (G,${\mu}$)), where G = (V,E) is a graph called the underlying graph of $S_n$ and ${\sigma}$:E ${\rightarrow}H_n({\mu}:V{\rightarrow}H_n)$ is a function. The restricted super line graph of index r of a graph G, denoted by $\mathcal{R}\mathcal{L}_r$(G). The vertices of $\mathcal{R}\mathcal{L}_r$(G) are the r-subsets of E(G) and two vertices P = ${p_1,p_2,{\ldots},p_r}$ and Q = ${q_1,q_2,{\ldots},q_r}$ are adjacent if there exists exactly one pair of edges, say $p_i$ and $q_j$, where $1{\leq}i$, $j{\leq}r$, that are adjacent edges in G. Analogously, one can define the restricted super line symmetric n-sigraph of index r of a symmetric n-sigraph $S_n$ = (G,${\sigma}$) as a symmetric n-sigraph $\mathcal{R}\mathcal{L}_r$($S_n$) = ($\mathcal{R}\mathcal{L}_r(G)$, ${\sigma}$'), where $\mathcal{R}\mathcal{L}_r(G)$ is the underlying graph of $\mathcal{R}\mathcal{L}_r(S_n)$, where for any edge PQ in $\mathcal{R}\mathcal{L}_r(S_n)$, ${\sigma}^{\prime}(PQ)$=${\sigma}(P){\sigma}(Q)$. It is shown that for any symmetric n-sigraph $S_n$, its $\mathcal{R}\mathcal{L}_r(S_n)$ is i-balanced and we offer a structural characterization of super line symmetric n-sigraphs of index r. Further, we characterize symmetric n-sigraphs $S_n$ for which $\mathcal{R}\mathcal{L}_r(S_n)$~$\mathcal{L}_r(S_n)$ and $$\mathcal{R}\mathcal{L}_r(S_n){\sim_=}\mathcal{L}_r(S_n)$$, where ~ and $$\sim_=$$ denotes switching equivalence and isomorphism and $\mathcal{R}\mathcal{L}_r(S_n)$ and $\mathcal{L}_r(S_n)$ are denotes the restricted super line symmetric n-sigraph of index r and super line symmetric n-sigraph of index r of $S_n$ respectively.