• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1285-1291
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• 2006

The proline utilization (put) operon of Vibrio vulnificus consists of the putAP genes encoding a proline dehydrogenase and proline permease. The result of put-lux transcriptional fusion analysis suggests that the V vulnificus putAP operon is not autoregulated by the PutA protein. A putR null mutation decreased proline dehydrogenase activity and the level of the put transcripts, indicating that transcription of putAP is under the positive control of PutR. The deduced amino acid sequence of the putR was similar to those reported from other bacteria with high levels of identity. Chromatin IP and GST pull-down assays revealed that PutR specifically binds to the putAP promoter region in vivo, and interacts with CRP in vitro. Taken together, the results suggested that PutR exerts its effect on putAP expression by directly interacting with CRP bound to the upstream region of P$_{put}$.

• 한국미생물·생명공학회지
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• 제19권6호
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• pp.568-574
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• 1991

효모에서 대랑의 물질생산계를 구축하기 위하여 먼저 여러 종류의 베터의 이용이 가능한 다양한 영양요구성 marker를 지니며 형질전환율이 향상된 효모숙주를 선별 개량하였다. 벡터의 제작에 사용되는 프로모터로는, 효모의 여러 유전자 중에서 그 활성이 매우 높은 해당계의 효소 GAP-DH의 구조 유전자 GAP를 이용하기로 하여, 효모염색체 DNA중에서 GAP 프로모터를 분리하여 이용하기 쉽게 변형하였다. 분리된 GAT promoter의 기능을 검토하기 위하여, reporter로 APase의 구조유전자 PHO5'를 이용하여 세포내의 copy수가 상이한 발현 벡터를 제작하여 GAP 프로모터에 의한 APase의 활성 및 전사산물을 측정한 결과, 정상적인 전사가 이루어 졌으며, 효소활성도 높게 나타났으며, 벡터의 copy수에 의한 효소활성의 차이도 감지되었다.

• Journal of Microbiology and Biotechnology
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• 제19권3호
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• pp.271-276
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• 2009

Functional gene studies in the cultivated white button mushroom Agaricus bisporus have been constrained by the absence of effective gene-silencing tools. Using two endogenous genes from A. bisporus, we have tested the utility of dsRNA hairpin constructs to mediate downregulation of specific genes. Hairpin constructs for genes encoding orotidine 5'-monophosphate decarboxylase (URA3) and carboxin resistance (CBX) were introduced into A. bisporus using Agrobacteriummediated transfection. Although predicted changes in phenotype were not observed in vitro, quantitative-PCR analyses indicated unambiguously that transcripts in several transformants were substantially reduced compared with the non-transformed controls. Interestingly, some hairpin transformants exhibited increased transcription of target genes. Our observations show that hairpin transgenic sequences can mediate downregulation of A. bisporus endogenous genes and that the technology has the potential to expedite functional genomics of the mushroom.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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• pp.1207-1215
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• 2014

Candida albicans, the major human fungal pathogen, undergoes morphological transition from the budding yeast form to filamentous growth in response to nitrogen starvation. In this study, we identified a new function of GST2, whose expression was required for filamentous growth of C. albicans under nitrogen-limiting conditions. The Gst2p showed Gst activity and required response to oxidative stress. The ${\Delta}gst2$ mutant displayed predominantly yeast phase growth in low ammonium media. Such morphological defect of ${\Delta}gst2$ mutants was not rescued by overexpression of Mep2p, Cph1p, or Efg1p, but was rescued by either overexpression of a hyperactive $RAS1^{G13V}$ allele or through exogenous addition of cyclic AMP. In addition, the ${\Delta}gst2$ mutants had lower levels of RAS1 transcripts than wild-type cells under conditions of nitrogen starvation. These results were consistent with the Ras1-cAMP pathway as a possible downstream target of Gst2p. These findings suggest that Gst2p is a significant component of nitrogen starvation-induced filamentation in C. albicans.

• Fisheries and Aquatic Sciences
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• 제12권2호
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• pp.90-97
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• 2009

The cytoskeletal $\beta$-actin gene and its 5'-upstream region were isolated and characterized in the rockbream (Oplegnathus fasciatus). Complementary DNA of the rockbream $\beta$-actin represented a 1,125 bp of an open reading frame encoding 375 amino acids, and the rockbream $\beta$-actin cDNA and deduced amino acid sequences were highly homologous to those of other vertebrate orthologs. At the genomic level, the $\beta$-actin gene also exhibited an organization typical of vertebrate cytoskeletal actin genes (2,159 bp composed of five translated exons interrupted by four introns) with a conserved GT/AG exon-intron splicing rule. The putative non-translated exon predicted in the rockbream $\beta$-actin gene was much more homologous with those of teleostean $\beta$-actin genes than those of mammals. The 5'-upstream regulatory region isolated by genome walking displayed conserved and essential elements such as TATA, CArG and CAAT boxes in its proximal part, while several other immune- or stress-related motifs such as those for NF-kappa B, USF, HNF, AP-1 and C/EBP were in the distal part. Semi-quantitative RT-PCR assay results demonstrated that the rockbream $\beta$-actin transcripts were ubiquitously but different-tially expressed across the tissues of juveniles.

• Fisheries and Aquatic Sciences
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• 제15권4호
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• pp.317-324
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• 2012

We characterized the cytoskeletal beta-actin (${\beta}$-ACT) gene (actb) and its 5'-upstream regulatory region in the Javanese ricefish Oryzias javanicus. The gene and protein structures were deduced from amino acid sequences of the actb gene and conserved in the teleost lineage. The O. javanicus actb gene has common transcription factor binding motifs in its regulatory region found in teleostean orthologues. Following quantitative reverse transcription-PCR, actb gene transcripts were detected in all tissues examined; however, the basal expression levels were different. During early development, O. javanicus actb mRNA levels showed a gradual increase and peaked between late somitogenesis and the heartbeat stage. Microinjection of O. javanicus embryos with the actb gene promoter-driven red fluorescent protein (RFP) gene reporter vector showed a ubiquitous distribution of RFP signals, although most exhibited a mosaic pattern of transgene expression. A small number of microinjected embryos displayed a wide distribution of RFP signals over their entire body, which resembled the expression pattern of endogenous actb. Data from this study provide a basis to develop a transgenic system with ubiquitous expression of foreign genes in O. javanicus.

• Fisheries and Aquatic Sciences
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• 제15권4호
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• pp.325-335
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• 2012

The present study characterized the fast skeletal myosin light chain-2 gene (mlc2f) in the euryhaline Javanese ricefish (Oryzias javanicus: Beloniformes). Coding nucleotide and deduced amino acid sequences of Javanese ricefish mlc2f were well conserved in the vertebrate lineage. Javanese ricefish mlc2f showed a typical seven-exon structure, and its promoter exhibited transcription factor binding motifs common to most muscle-specific genes. However, Javanese ricefish mlc2f also displayed tandem duplications of intronic sequences in both intron 1 and intron 3. Based on quantitative reverse transcription-polymerase chain reaction, the mlc2f transcripts were highly predominant in skeletal muscles of adults and were differentially modulated during embryonic development. Microinjection of the mlc2f promoter-driven red fluorescent protein (RFP) reporter construct successfully exhibited heterologous expression of the fluorescent reporter, primarily in muscular areas of hatchlings, although the distribution pattern of RFP signals was not uniform due to the mosaic nature of the introduced transgene. Data from this study indicate that the Javanese ricefish mlc2f gene has undergone "intra-intronic" duplication events in a species-specific manner and that the mlc2f regulator may also be useful in heterologous expression assays of the skeletal muscles of this species.

• International Journal of Industrial Entomology and Biomaterials
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• 제10권1호
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• pp.73-77
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• 2005

A Cu,Zn superoxide dismutase (SOD1) cDNA was cloned from the spider, Araneus ventricosus. The A. ventricosus SOD1 (AvSOD1) cDNA contains an open reading frame of 495 bp encoding 165 amino acid polypeptide with a predicted molecular mass of 17,114 Da and pI of 6.55, and possesses the typical metal binding ligands of six histidines and one aspartic acid common to SOD1s. The deduced amino acid sequence of the AvSOD1 cDNA showed $51\%$ identity to Ceratitis capitata SOD1, and $50\%$ to SOD1 sequences of both Drosophila melanogaster and Chymomyza amoena. Northern blot analysis revealed the presence of AvSOD1 transcripts in all tissues examined.

• 대한간호학회지
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• 제47권1호
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• pp.110-120
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• 2017

Purpose: The purpose of this study was to describe the lived experience of patients with heart transplantation in Korea. Methods: Individual indepth interviews and a focus group interview were used to collect the data from nine patients who had heart transplantations in 2015. All interviews were audio-taped and verbatim transcripts were made for the analysis. Data were analyzed using Colaizzi's phenomenological method. Results: Among the nine participants, eight were men. Mean age was 57.30 years. Six theme clusters emerged from the analysis. 'Joy of rebirth obtained by good luck' describes the pleasure and expectation of new life after narrow survival. 'Suffering from adverse drug effects' illustrates various psychosocial difficulties, such as low self-esteem, helplessness, alienation, and burnout, arising from the side effects of medications. 'Body and mind of being bewildered' illustrates disintegrated health and haunting fear of death. 'Alienation disconnected with society' describes isolated feeling of existence due to misunderstandings from society. 'Suffering overcome with gratitude and responsibility' includes overcoming experience through various social supports and suitable jobs. Finally, 'acceptance of suffering accompanied with new heart' illustrate changed perspective of life itself. Conclusion: The findings in this study provide deep understanding and insights of the lived experience of heart related illness for these patients and should help in the development of tailored-interventions for patients with heart transplantation.

• 한국직업건강간호학회지
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• 제20권2호
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• pp.185-194
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• 2011

Purpose: Global shortages in nursing and strong demand for nursing services provided Korean nurses with more overseas employment opportunities, especially in the developed countries such as the United States and Canada. The purpose of this study was to explore the ageing experiences of the Korean nurses dispatched to Germany in the 1960s and 1970s. Methods: The researcher interviewed 10 Korean nurses living in four cities in Germany. Interviews were performed twice in January and July, 2010, and the data, in the form of field notes and interview transcripts, were analyzed using the Agar's (1980) ethnographic method. Results: The ageing experiences of the participants can be summarized into three theme stages: coming upon old age, reluctantly realizing getting old, and finally accepting being old. The first stage is characterized by 'wiring money to homeland all throughout youth', second 'still feeling like a stranger anywhere', and finally 'burying homeland in heart'. Conclusion: The research findings not only suggest crucial materials for training prospective nurses overseas for their successful settlement, but also shed lights on related problems and solutions with ageing experience in overseas employment.