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PCR-Based Sensitive Detection and Identification of Xanthomonas oryzae pv. oryzae (중합효소연쇄 반응에 의한 벼 흰잎마름병균의 특이적 검출)

  • Lee, Byoung-Moo;Park, Young-Jin;Park, Dong-Suk;Kim, Jeong-Gu;Kang, Hee-Wan;Noh, Tae-Hwan;Lee, Gil-Bok;Ahn, Joung-Kuk
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.256-264
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    • 2004
  • A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.

Introduction of Bean Chitinase Gene into Korean Ginseng by Agrobaterium tumefaciens (Agrobacterium tumefaciens에 의한 강낭콩 키틴가수분해효소 유전자의 고려인삼으로의 도입)

  • 이행순;권석윤;백경희;김석원;이광웅;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.95-99
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    • 1995
  • We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

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A Study on the Web-based Map Algebraic Processor (웹 기반 지도대수 처리기에 관한 연구)

  • 박기호
    • Spatial Information Research
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    • v.5 no.2
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    • pp.147-160
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    • 1997
  • "The "Map Algebra", beeing recognized as a viable theoretical framework for GIS (Geographica Infonnation System), models map layers as "operands" which are the basic unit of geo-processing, and a variety of GIS commands as "operators." In this paper, we attempt at lifting some limitations of map algebras proposed in GIS literature. First, we model map layer as "function" such that we may employ the notion of meta operator (or, higher-order funtion) available in the functional programming paradigm. This approach provides map algebraic language with "programmability" needed in GIS user language. Second, we extend the semantics of, and improve on the sytactic structure of map algebraic language. Mer the data model and language associated with map algebra are formalized, we proceed to design and implement a prototype of map algebraic processor. The parser of the language in our prototype plays the role of transforming the native and heterogeneous user language of current GISs into a canonical map algebraic language. The prototype, named "MapSee" is a proof-of-concept system for the ideas we propsed in this paper. We believe that the uniform interface based on the map algebraic language will make promising infrastructure to support "Internet GIS." This is because the uniform but powerful interface through the Web clients allow access to both geo-data and geo-processing resources distributed over the network.to both geo-data and geo-processing resources distributed over the network.

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Fine Mapping of the Rice Bph1 Gene, which Confers Resistance to the Brown Planthopper (Nilaparvata lugens Stal), and Development of STS Markers for Marker-assisted Selection

  • Cha, Young-Soon;Ji, Hyeonso;Yun, Doh-Won;Ahn, Byoung-Ohg;Lee, Myung Chul;Suh, Seok-Cheol;Lee, Chun Seok;Ahn, Eok Keun;Jeon, Yong-Hee;Jin, Il-Doo;Sohn, Jae-Keun;Koh, Hee-Jong;Eun, Moo-Young
    • Molecules and Cells
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    • v.26 no.2
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    • pp.146-151
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    • 2008
  • The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.

The Effects of MERS(Middle East Respiratory Syndrome) Event on the Psychosocial Wellbeing of Healthcare Workers and the Public with the Mediating Effect of Resilience (메르스(Middle East Respiratory Syndrome)사태에 따른 병원종사자와 일반인의 스트레스 정도가 사회심리적 건강에 미치는 영향과 회복탄력성의 매개효과)

  • Kwon, Hyuk-Min;Kim, Tae-Hyung;Choi, Mal-Rye;Kim, Byoung-Jo;Kim, Hyoung-Wook;Song, Ok-Sun;Eun, Hun-Jeong
    • Korean Journal of Psychosomatic Medicine
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    • v.25 no.2
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    • pp.111-119
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    • 2017
  • Objectives : The purpose of this study is to investigate the effects of stress on psychosocial wellbeing at the time of an outbreak of Middle East respiratory syndrome(MERS) and to investigate the effect of resilience as a mediator on the relationship between stress and psychosocial wellbeing. Methods : Perceived Stress Scale, Psychosocial Wellbeing Index Short Form, and the Conner-Davidson Resilience Scale was implemented for 156 medical persons who worked at the hospital in which exposure to MERS cases had been confirmed and 127 ordinary people. We conducted a Pearson correlation coefficient and a hierarchical multiple regression to confirm the effect of stress on psychosocial wellbeing and the mediating effect of resilience between stress and psychosocial wellbeing. Results : The higher the perceived stress, the lower the psychosocial wellbeing in both healthcare workers and the public. The higher the perceived stress, the lower the resilience and the research results showed that there was a partially mediating effect of resilience in the relationship between stress and psychosocial wellbeing. Conclusions : This study demonstrated that the degree of individual resilience can indirectly give a positive effect on the psychosocial wellbeing when people under the stress by MERS shows adverse effects on psychosocial wellbeing. This suggests that clinical intervention and psychosocial approach aiming at strengthening resilience is important to maintain mental health during crisis development.

Epidemiology, Virology, and Clinical Features of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2; Coronavirus Disease-19) (코로나바이러스감염증-19의 바이러스 (SARS-CoV-2) 특징, 전파 및 임상 양상)

  • Park, Su Eun
    • Pediatric Infection and Vaccine
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    • v.27 no.1
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    • pp.1-10
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    • 2020
  • A cluster of severe pneumonia of unknown etiology in Wuhan City, Hubei province in China emerged in December 2019. A novel coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was isolated from lower respiratory tract sample as the causative agent. The current outbreak of infections with SARS-CoV-2 is termed coronavirus disease 2019 (COVID-19) by the World Health Organization (WHO). COVID-19 rapidly spread into at least 114 countries and killed more than 4,000 people by March 11, 2020. WHO officially declared COVID-19 a pandemic on March 11, 2020. There have been 2 novel coronavirus outbreaks in the past 2 decades. The outbreak of severe acute respiratory syndrome (SARS) in 2002-2003 caused by SARS-CoV had a case fatality rate of around 10% (8,098 confirmed cases and 774 deaths), while Middle East respiratory syndrome (MERS) caused by MERS-CoV killed 858 people out of a total 2,499 confirmed cases between 2012 and 2019. The purpose of this review is to summarize known-to-date information about SARS-CoV-2, transmission of SARS-CoV-2, and clinical features of COVID-19.

Mutation Analysis of Synthetic DNA Barcodes in a Fission Yeast Gene Deletion Library by Sanger Sequencing

  • Lee, Minho;Choi, Shin-Jung;Han, Sangjo;Nam, Miyoung;Kim, Dongsup;Kim, Dong-Uk;Hoe, Kwang-Lae
    • Genomics & Informatics
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    • v.16 no.2
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    • pp.22-29
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    • 2018
  • Incorporation of unique barcodes into fission yeast gene deletion collections has enabled the identification of gene functions by growth fitness analysis. For fine tuning, it is important to examine barcode sequences, because mutations arise during strain construction. Out of 8,708 barcodes (4,354 strains) covering 88.5% of all 4,919 open reading frames, 7,734 barcodes (88.8%) were validated as high-fidelity to be inserted at the correct positions by Sanger sequencing. Sequence examination of the 7,734 high-fidelity barcodes revealed that 1,039 barcodes (13.4%) deviated from the original design. In total, 1,284 mutations (mutation rate of 16.6%) exist within the 1,039 mutated barcodes, which is comparable to budding yeast (18%). When the type of mutation was considered, substitutions accounted for 845 mutations (10.9%), deletions accounted for 319 mutations (4.1%), and insertions accounted for 121 mutations (1.6%). Peculiarly, the frequency of substitutions (67.6%) was unexpectedly higher than in budding yeast (~28%) and well above the predicted error of Sanger sequencing (~2%), which might have arisen during the solid-phase oligonucleotide synthesis and PCR amplification of the barcodes during strain construction. When the mutation rate was analyzed by position within 20-mer barcodes using the 1,284 mutations from the 7,734 sequenced barcodes, there was no significant difference between up-tags and down-tags at a given position. The mutation frequency at a given position was similar at most positions, ranging from 0.4% (32/7,734) to 1.1% (82/7,734), except at position 1, which was highest (3.1%), as in budding yeast. Together, well-defined barcode sequences, combined with the next-generation sequencing platform, promise to make the fission yeast gene deletion library a powerful tool for understanding gene function.

Phylogenetic relationships of genera Trametes on the basis of ITS region sequences (rDNA의 ITS 부위 염기서열 분석에 의한 구름버섯 균주의 유전적인 유연관계 분석)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Oh, Jin-A;Han, Hye-Su;Um, Na-Na
    • Journal of Mushroom
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    • v.9 no.1
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    • pp.27-33
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    • 2011
  • This study was carried to identify a correct species and asses genetic diversity within the same species of Trametes spp. preserved in Division of applied Microbiology The morphological and cultural characteristics of preserved strains were observed through microscope and investigated on PDA, respectively. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Trametes spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that five strains were different species and six strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Trametes clustered into four distinct group, most of which correlated with species-groups identified by RAPD method. Seven isolates included TM 01 strain showed high similarity with Trametes versicolr, TM 07 and TM 10 high similarity with Trametes gibbosa, and TM 05 high similarity with Trametes elegans. But isolates collected in the United States was identified as T. junipericola. T. gibbosa and T. versicolor by RAPD analysis of genetic polymorphisms showed a very different band patterns and these strains showed different band patterns on areas. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 11 isolates of Trametes spp. may be need to reclassify or eliminate from preserved catalogue.

Development of a psychrophilic-SCAR marker for Pleurotus eryngii (큰느타리버섯의 저온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Chul;Hwang, Hye Sung;Cho, Yun Jun;Kim, Hye Su;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.171-176
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    • 2013
  • Genomic DNAs of psychrophilic strains of Pleurotus eryngii were analyzed by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S3 primers to develop the strain-specific DNA marker. A unique DNA fragment with the size of 480 bp was yielded by OP-S3 primer from the psychrophilic strain. A sequence characterized amplified region (SCAR) marker, designated as OP-S3-1, was designed on the basis of the determined sequence. The PCR analysis with the OP-S3-1 primer showed that this SCAR marker can clearly distinguish the psychrophilic strains from the control strains.

Discrimination of Potato Varieties by Random Amplified Polymorphic DNA Analysis (RAPD에 의한 감자 품종의 구분)

  • Seo, Hyo Won;Yi, Jung Yoon;Cho, Hyun Mook;Park, Young Eun;Oh, Seung Eun
    • Horticultural Science & Technology
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    • v.19 no.1
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    • pp.29-33
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    • 2001
  • This study was carried out to discriminate potato cultivars and breeding lines by specific molecular markers using random amplified polymorphic DNA (RAPD) analysis. The genotypes of potatoes used for analysis were eight cultivars and five breeding lines. Some of those show much phenotypic resemblances among them because 'Jopung', 'Daekwan70', 'Gawon', and 'Daekwan72' have immediate parental relationship with 'Superior', 'Irish Cobbler', 'Namsuh', and 'Atlantic', respectively. So, there are many difficulties to distinguish the varieties by the morphological characteristics. Three URP primers, URP2, URP4, and URP8 were selected for promising primers to discriminate potato genotypes or cultivars. The three URP primers were shown very high reproducibility because of the relatively high annealing temperature and long primer size. Although the results of similarity analyses did not always reflect the genetic relationship between potato varieties, the reproducible pattern of amplified DNA bands by URP primers showed possibility for molecular markers for discrimination of potato genotype or cultivar.

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