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Introduction of Shiva Gene into tobacco and Potato Using Tissue-Specific Tomato PAL Promoter (조직특이성 promoter를 이용한 Shiva 유전자의 식물체내 도입)

  • 이정윤;이신우;박권우
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.109-113
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    • 1998
  • In this study we tried to transform an antimicrobial peptide gene (Shiva) under the promoter of tomato phenylalanine ammonia-lyase (tPAL5) into tobacco and potato plants. Antimicrobial peptide gene was isolated originally from giant silk moth (Hyalophora cecropia) and modified ie nucleotide sequence to increase antimicrobial activity. Transgenic tobacco plants were regenerated and their seeds were tested on the media containing kanamycin (500 mg/L). The results of PCR amplification and genomic Southern blot hybridization confirmed the integration of construct (tPAL5 promoter-Shiva-NOS-GUS-NOS) into chromosome. We observed that one of the transgenic tobacco plants showed chromosome rearrangement when integrated. In case of potato transformation, the efficiency of regeneration was maximized at the medium containing Zeatin 2mg/L, NAA 0.01mg/L, GA$_3$ 0.1mg/L. We also observed the high expression of GUS (${\beta}$-glucuronidase) enzyme which was located next to the terminator sequence of nopaline synthase gene (NOS) in the vascular tissue of stem, leaves of transgenic potatoes. This result suggested that a short sequence of Shiva gene (120 bp) and NOS terminator sequence might be served as a leader sequence of transcript when translated.

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Up-regulation of dynamin-2 gene expression in Ras-transformed cells (Ras에 의해 암화된 세포에서 dynamin-2의 발현 촉진)

  • Yoo, Ji-Yun
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.375-380
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    • 2007
  • Dynamin plays a key role in the scission event common to various types of endocytosis. It has been previously reported that the SH3 domain-mediated association of Grb2 with dynamin-2 was dominantly found in Ras transformed cells. However, whether this association results from the increased expression of dynamin-2 and Grb2 in Ras transformed cells or not is still unknown. So in this study we first analyzed the expression levels of dynamin-2 and Grb2 and found that the expression of dynamin-2 protein was dramatically increased in Ras-transformed NIH3T3 (NIH3T3(Ras)) cells. Furthermore competitive PCR data revealed that the mRNA transcripts for dynamin-2 were increased about 100-fold in NIH3T3(Ras) compared to those of NIH3T3 cells. However, the protein level and mRNA transcript of Grb2 were not changed in these two cells. We also examined promoter activity of dynamin-2 in NIH3T3(Ras) cells and suggest the existence of Ras-responsive sequence in promoter region -300 to -200.

Phylogenetic Study of Genus Sorbus in Korea by Internal Transcribed Spacer Sequence (ITS) (ITS에 의한 한국내 마가목 속 분류군의 유전적 계통분류학적 연구)

  • Huh, Man-Kyu;Kim, Sea-Hyun;Park, So-Hye
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1610-1615
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    • 2007
  • Genus Sorbus is a long lived woody species that is primarily distributed throughout Asia and Europe. This species is regarded as very important herbal medicines in Korea and China. Sorbus commixta is primarily distributed throughout Europe. We evaluated a representative sample of the four taxa with nuclear ribosomal DNA internal transcribed spacer sequences (ITS) to estimate genetic relationships within genus. Aligned nucleotide sequences of the length of ITS1 were nearly constant within genus Sorbus varying from 219 in S. aucuparia to 218 in the rest species. Especially, the 5.8S subunit of all taxa of Sorbus was found to constant of 165 bp nucleotides. However, aligned nucleotide sequences of the length of ITS2 vary from 240 in S. sambucifolia var. pseudogrcilisto 245 in S. aucuparia. Total alignment length is 629 positions, of which 35 are parsimony-informative, 32 variable but parsimony-uninformative, and 552 constant characters. The base furtherance showed the difference to the by a total taxon: an average A and T are 17.7% and G and C are 30.4%, 34.2%, respectively. All the four taxa beginning with conserved base paired triplets emerging from single strand regions (domain I). Noteworthy, in the RNA secondary structure proposed for the three Korean Sorbus taxa RNA transcript ITS2, which shows a remarkedly well-conserved folding (domain II). When compared to the European Sorbus (S. aucuparia) of ITS2. ITS analysis may be useful in germ-plasm classification several taxa of genus Sorbus.

Rice OsACDR1 (Oryza sativa Accelerated Cell Death and Resistance 1) Is a Potential Positive Regulator of Fungal Disease Resistance

  • Kim, Jung-A;Cho, Kyoungwon;Singh, Raksha;Jung, Young-Ho;Jeong, Seung-Hee;Kim, So-Hee;Lee, Jae-eun;Cho, Yoon-Seong;Agrawal, Ganesh K.;Rakwal, Randeep;Tamogami, Shigeru;Kersten, Birgit;Jeon, Jong-Seong;An, Gynheung;Jwa, Nam-Soo
    • Molecules and Cells
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    • v.28 no.5
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    • pp.431-439
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    • 2009
  • Rice Oryza sativa accelerated cell death and resistance 1 (OsACDR1) encodes a putative Raf-like mitogen-activated protein kinase kinase kinase (MAPKKK). We had previously reported upregulation of the OsACDR1 transcript by a range of environmental stimuli involved in eliciting defense-related pathways. Here we apply biochemical, gain and loss-of-function approaches to characterize OsACDR1 function in rice. The OsACDR1 protein showed autophosphorylation and possessed kinase activity. Rice plants overexpressing OsACDR1 exhibited spontaneous hypersensitive response (HR)-like lesions on leaves, upregulation of defense-related marker genes and accumulation of phenolic compounds and secondary metabolites (phytoalexins). These transgenic plants also acquired enhanced resistance to a fungal pathogen (Magnaporthe grisea) and showed inhibition of appressorial penetration on the leaf surface. In contrast, loss-of-function and RNA silenced OsACDR1 rice mutant plants showed downregulation of defense-related marker genes expressions and susceptibility to M. grisea. Furthermore, transient expression of an OsACDR1:GFP fusion protein in rice protoplast and onion epidermal cells revealed its localization to the nucleus. These results indicate that OsACDR1 plays an important role in the positive regulation of disease resistance in rice.

Systematic Review of Short Stature and Growth Related Qualitative Researches (저신장 및 성장과 관련된 질적연구에 대한 체계적 문헌 고찰)

  • Kim, Hye Jin;Jeon, Chae Heun;Roh, Min Yeong;Han, Ye Ji;Kim, Hyun Ho;Cheong, Moon Joo;Leem, Jung Tae
    • The Journal of Pediatrics of Korean Medicine
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    • v.33 no.4
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    • pp.74-88
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    • 2019
  • Objectives The aim of this study is systematic evaluation of the quality of qualitative studies related to short stature. It will be utilized as fundamental data for future qualitative research about Korean Medicine Pediatrics treatment for short stature. Methods We searched qualitative studies related to short stature in the Pubmed. Two researchers independently assessed the quality of the qualitative researches by using Consolidated criteria for reporting qualitative research (COREQ) and Critical Appraisal Skills Program (CASP). We also qualitatively synthesized theme of each included studies. We presented integrated theme and categories with in vivo coding about qualitative research about short stature. Results A total of 47 papers were screened and 7 papers were finally selected. In the quality assessment using COREQ, detalied information about the researchers, methods of approach, reasons for non-participation, presence of non-participants, participant's review of the transcript, and participant's review of the findings were insufficient. In the quality evaluation using CASP, there were lack of justifications for the research methods, description method of research, and bias in process of data analysis. The findings were synthesized into three themes: fixed idea about height, discomfort caused by small stature and personal coping strategy with stress caused by short stature. Conclusions In further qualitative research, it is necessary to report the research according to COREQ and CASP checklist. It is also necessary to understand the stress and coping strategies to short stature of patients, We need to develop a social management strategy for short stature.

mRNA Expression of the Regulatory Factors for the Early Folliculogenesis in vitro (체외배양 중인 생쥐 난소에서 초기난포 조절인자의 발현)

  • Yoon, Se-Jin;Kim, Ki-Ryeong;Chung, Hyung-Min;Yoon, Tae-Ki;Cha, Kwang-Yul;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.3
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    • pp.207-216
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    • 2005
  • Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries. Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in ${\alpha}$-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control. Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro. Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.

Comparison of Digital PCR and Quantitative PCR with Various SARS-CoV-2 Primer-Probe Sets

  • Park, Changwoo;Lee, Jina;Hassan, Zohaib ul;Ku, Keun Bon;Kim, Seong-Jun;Kim, Hong Gi;Park, Edmond Changkyun;Park, Gun-Soo;Park, Daeui;Baek, Seung-Hwa;Park, Dongju;Lee, Jihye;Jeon, Sangeun;Kim, Seungtaek;Lee, Chang-Seop;Yoo, Hee Min;Kim, Seil
    • Journal of Microbiology and Biotechnology
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    • v.31 no.3
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    • pp.358-367
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    • 2021
  • The World Health Organization (WHO) has declared the coronavirus disease 2019 (COVID-19) as an international health emergency. Current diagnostic tests are based on the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method, which is the gold standard test that involves the amplification of viral RNA. However, the RT-qPCR assay has limitations in terms of sensitivity and quantification. In this study, we tested both qPCR and droplet digital PCR (ddPCR) to detect low amounts of viral RNA. The cycle threshold (CT) of the viral RNA by RT-PCR significantly varied according to the sequences of the primer and probe sets with in vitro transcript (IVT) RNA or viral RNA as templates, whereas the copy number of the viral RNA by ddPCR was effectively quantified with IVT RNA, cultured viral RNA, and RNA from clinical samples. Furthermore, the clinical samples were assayed via both methods, and the sensitivity of the ddPCR was determined to be equal to or more than that of the RT-qPCR. However, the ddPCR assay is more suitable for determining the copy number of reference materials. These findings suggest that the qPCR assay with the ddPCR defined reference materials could be used as a highly sensitive and compatible diagnostic method for viral RNA detection.

A Study on the Open Meeting System : Focused on the Central Administrative Organization Designated Meeting (회의공개제도 운영에 관한 연구 중앙행정기관 지정회의를 중심으로)

  • Choi, Jeong Min;Kim, You-seung
    • The Korean Journal of Archival Studies
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    • no.55
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    • pp.307-334
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    • 2018
  • This study analyzes the status of the designated administrative meetings of the central administrative agencies, including the number of meetings, the presence of written minutes, and the form of minutes. The implications based on the analysis are as follows. First, there were meetings that were not held, and written meetings were held. The standards for determining the designated meetings should be established. Second, when only a written meeting was held, the minutes were not made because these were not written separately. A written meeting can be held, if it is necessary. However, there is still a need to keep a detailed record similar to minutes in such cases. Third, among the minute types, a written record of the points of the meeting is preferred. Even if the meeting is closed for proper reasons, making a transcript and a record in addition to writing the points must be settled. Finally, specific standards for the procedures and reasons why meetings and minutes should be closed should be established. It is necessary to change the paradigm from the viewpoint of production and management to the viewpoint of disclosure and sharing through the improvement of standards and procedures related to designated meetings and designated meeting minutes.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat

  • Xie, Li-hua;Quan, Xin;Zhang, Jie;Yang, Yan-yan;Sun, Run-hong;Xia, Ming-cong;Xue, Bao-guo;Wu, Chao;Han, Xiao-yun;Xue, Ya-nan;Yang, Li-rong
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.11-18
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    • 2019
  • Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. $TUB{\beta}$ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of $TUB{\beta}$ expression. The expression profile of ExoPG assessed using $TUB{\beta}$ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

A Classification Model for Illegal Debt Collection Using Rule and Machine Learning Based Methods

  • Kim, Tae-Ho;Lim, Jong-In
    • Journal of the Korea Society of Computer and Information
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    • v.26 no.4
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    • pp.93-103
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    • 2021
  • Despite the efforts of financial authorities in conducting the direct management and supervision of collection agents and bond-collecting guideline, the illegal and unfair collection of debts still exist. To effectively prevent such illegal and unfair debt collection activities, we need a method for strengthening the monitoring of illegal collection activities even with little manpower using technologies such as unstructured data machine learning. In this study, we propose a classification model for illegal debt collection that combine machine learning such as Support Vector Machine (SVM) with a rule-based technique that obtains the collection transcript of loan companies and converts them into text data to identify illegal activities. Moreover, the study also compares how accurate identification was made in accordance with the machine learning algorithm. The study shows that a case of using the combination of the rule-based illegal rules and machine learning for classification has higher accuracy than the classification model of the previous study that applied only machine learning. This study is the first attempt to classify illegalities by combining rule-based illegal detection rules with machine learning. If further research will be conducted to improve the model's completeness, it will greatly contribute in preventing consumer damage from illegal debt collection activities.