• Title/Summary/Keyword: 1-sequence covering

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Optimization of Polyethylene Glycol-Mediated Transformation of the Pepper Anthracnose Pathogen Colletotrichum scovillei to Develop an Applied Genomics Approach

  • Shin, Jong-Hwan;Han, Joon-Hee;Park, Hyun-Hoo;Fu, Teng;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.575-584
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    • 2019
  • Colletotrichum acutatum is a species complex responsible for anthracnose disease in a wide range of host plants. Strain C. acutatum KC05, which was previously isolated from an infected pepper in Gangwon Province of South Korea, was reidentified as C. scovillei using combined sequence analyses of multiple genes. As a prerequisite for understanding the pathogenic development of the pepper anthracnose pathogen, we optimized the transformation system of C. scovillei KC05. Protoplast generation from young hyphae of KC05 was optimal in an enzymatic digestion using a combined treatment of 2% lysing enzyme and 0.8% driselase in 1 M NH4Cl for 3 h incubation. Prolonged incubation for more than 3 h decreased protoplast yields. Protoplast growth of KC05 was completely inhibited for 4 days on regeneration media containing 200 ㎍/ml hygromycin B, indicating the viability of this antibiotic as a selection marker. To evaluate transformation efficiency, we tested polyethylene glycol-mediated protoplast transformation of KC05 using 19 different loci found throughout 10 (of 27) scaffolds, covering approximately 84.1% of the entire genome. PCR screening showed that the average transformation efficiency was about 17.1% per 100 colonies. Southern blot analyses revealed that at least one transformant per locus had single copy integration of PCR-screened positive transformants. Our results provide valuable information for a functional genomics approach to the pepper anthracnose pathogen C. scovillei.

Test Case Generation of Communication Protocol with Regular Expressions (정규표현식을 이용한 통신 프로토콜의 최소 시험 경로 생성)

  • 김한경
    • Journal of Internet Computing and Services
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    • v.2 no.1
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    • pp.1-11
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    • 2001
  • Though it is proposed to use Petri net or dynamic FSM methods for the generation of test sequences on some specific protocols, those methods ere unavailable on the cases where the protocol allows faults processing or includes paths in looping which cause errors or endless looping by the explosion of states. The determination of test coverage on the protocol software that has been designed and implemented is difficult by the reason of development periods, technical solutions to support and also economical limitations. It is suggested to generate timely protocol software test sequences on the basis of regular expressions covering the functions of protocol. With this regular expression method, the 38 test sequences of Q.2971 protocol has been generated and also minimized the endless looping problem when dynamic test suites are used by simplifying the test path expressions that denotes loops, According to the works, the suggested method is confirmed as simple and easy compare to the other dynamic test sequence generation techniques. Moreover. the method to search an optional test path whether it Is included or not in the regular path expression is reviewed.

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Construction of Complementary DNA Library and cDNA Cloning for Cy Strain of Odontoglossum Ringspot Virus Genomic RNA (오돈토글로썸 윤문 바이러스 Cy계통 게놈 RNA의 cDNA 구축 및 유전자 크로닝)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.228-234
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    • 1994
  • Genomic RNA was extracted from Cy strain of odontoglossum ringspot tobamovirus (ORSV-Cy) isolated from infected leaves of tobacco cv. Samsun. Size of the genomic RNA was about 6.6 kb in length. The genomic RNA was fractionated using Sephadex G-50 column chromatography into 2 fractions. They were polyadenylated at their 3'-end using E. coli poly(A) polymerase. Polyadenylated viral RNA was recovered by oligo (dT) primer adapter containing NotI restriction site and Moloney murine leukemia virus SuperScript reverse transcriptase (RNase H-). Second-strand cDNA was synthesized by using E. coli DNA ligase, E. coli DNA polymerase I and E. coli RNase H. Recombinant plasmids containing cDNAs for ORSV-Cy RNA ranged from about 800 bp to 3,000 bp. Among the selected 238 recombinants, pORCY-124 clone was the largest one covering 3'-terminal half of the viral RNA. This clone contained two restriction sites for EcoRI and XbaI and one site for AccI, AvaI, BglII, BstXI, HindIII, PstI, and TthIII 1. respectively. The clone contained partial viral replicase, a full-length movement protein and a complete coat protein genes followed by a 3' untranslated region of 414 nucleotides based on restriction mapping and nucleotide sequencing analyses. Clones pORCY-028, -068, -072, -187 and -224 were overlapped with the pORCY-124. Clones pORCY-014 and -095 covered 5' half upstream from the middle region of the viral RNA, which was estimated based on restriction mapping and partial sequence analysis. Constructed cDNA library covered more than 90% of the viral genome.

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A Study on the Sequence of Contents in Clothing and textile Education (의생활 교육내용의 계열화에 관한 연구)

  • 박일록;이은희
    • Journal of Korean Home Economics Education Association
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    • v.14 no.1
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    • pp.39-54
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    • 2002
  • This study was done to identify contents of clothing and textile education and to systemize its curriculum from elementary school to high school Every time the official curriculum is updated. the suggestion that the contents of Home Economics Education should be systematized never fails to come forth. Unfortunately. it's a fact that there are still insufficient grounds to base this opinion on. and that adequate research has not yet been done in this particular field. Therefore. basic research must be performed in order to systematize the substance of clothing and textile education. Keeping this in mind. the following study strives to find appropriate methods of conducting clothing and textile education, while attempting to provide basic materials for future organization and selection of contents. In order to achieve these goals. the materials have been sorted into three categories : First of all. what sort of concepts is being taught in the subject of clothing and textile. and are the chapters constructed so that different schools can maintain consistency among them\ulcorner Second, how has clothing and textile education been done so far and is the contents correctly synchronized with the students'development\ulcorner Third. haw is clothing and textile education utilized in real life. and has it actually been systematized in a way that encourages practical utilization\ulcorner This study has mostly been done through the research of various documental resources. Also. the analysis has been performed using the systematic contents analysis method. The main results of obtained this study are as follows : As the concepts of clothing and textile education. which are different from each school, has not completely been systematizied. Also. as the contents of clothing and textile education has not correctly synchronized with the students'development. am it has not actually been systematized in a way that encourages practical utilization. Since this study has touched only limited research on the topic of clothing and textile education. it is essential that further research covering a wider array of themes be done in the future.

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Identification of QTLs Associated with Resistance to Riptortus clavatus Thunberg (Heteroptera: Alydidae) in Soybean (Glycine max L. Merr.)

  • Li, Wenxin;Van, Kyujung;Zheng, Da-Hao;Liu, Weixian;Lee, Yeong-Ho;Lee, Sue-Yeon;Lee, Joon-Ho;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.11 no.4
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    • pp.243-248
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    • 2008
  • The bean bug Riptortus clavatus Thunberg (Heteroptera: Alydidae) is an important pest, causing serious yield loss in soybean. But the information on mechanism of resistance to R. clavatus is limited. The objective of this study was to identify QTLs for R. clavatus resistance using simple sequence repeat (SSR) markers in a soybean population of recombinant inbred lines (RILs) developed from the cross PI 171451 ${\times}$ Hwaeomputkong. A genetic map from this population was constructed with a total of 136 SSR markers covering 1073.9 cM on 20 linkage groups (LGs). With 126 $F_5$ RILs, two independent QTLs for resistance to R. clavatus were mapped on LGs B1 and C2. The amount of phenotypic variation explained by these QTLs ranged from 12 to 16%. PI 171451 showed an escape response to R. clavatus. Under feeding conditions, 14.4% of RILs showed greater resistance to R. clavatus than the resistant parent. The resistance to R. clavatus in soybean from PI 171451 was incomplete and quantitatively inherited and the QTLs for resistance to R. clavatus detected in the RIL population were not significantly affected by epistatic interactions.

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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.

A Study on the Quantitative Analysis for the Forest Landscape (삼림경관에 관한 계량적 분석에 관한 연구)

  • 서주환
    • Journal of the Korean Institute of Landscape Architecture
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    • v.15 no.1
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    • pp.39-67
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    • 1987
  • The purpose of this thesis is to suggest objective basic data for the environmental design through the quantitative analysis of the visual quality included in the physical environment of forest landscape. For this, landscape values of forest landscape have been evaluated by using the Iverson method, the images structure of forest landscape's main utilizing space have been analysed by the factor analysis algorithm, degree of visual preferences have been pleasured mainly by questionnaries and SBE method, and finally these thesis can be summarized as fallow LCP with high values of Iverson factors I and IV yield high landscape value. Specifically, Iverson factor IV has been found to play the dominant. For all experimental points, significant seasonal variations in S.D. scale values have been observed. In natural parks, where artificial structures are complementary to the natural landscape, main factors of image are S.D. scales such as the visual sequence, the formal simplicity of structures, the emphasis, the unification of heterogeneous factors and the assimilation. Factors covering the spatial image of natural parks have been found to be the overall evaluation, the individual characteristics, the tidiness, the potentiality, the dignity, the intimacy and the space volume. For all seasons, factors such as the individual characteristics, the dignity, the tidiness, the potentiality, yield high factor scores. As for factors determining the degree of visual preference, variables such as the summit, the skyline, rocks, the water and the degree of natural destruction by artificial structures yield high values for all seasons.

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Neural correlations of familiar and Unfamiliar face recognition by using Event Related fMRI

  • Kim, Jeong-Seok;Jeun, Sin-Soo;Kim, Bum-Soo;Choe, Bo-Young;Lee, Hyoung-Koo;Suh, Tae-Suk
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2003.09a
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    • pp.78-78
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    • 2003
  • Purpose: This event related fMRI study was to further our understanding about how different brain regions could contribute to effective access of specific information stored in long term memory. This experiment has allowed us to determine the brain regions involved in recognition of familiar faces among non familiar faces. Materials and Methods: Twelve right handed normal, healthy volunteer adults participated in face recognition experiment. The paradigm consists of two 40 familiar faces, 40 unfamiliar faces and control base with scrambled faces in a randomized order, with null events. Volunteers were instructed to press on one of two possible buttons of a response box to indicate whether a face was familiar or not. Incorrect answers were ignored. A 1.5T MRI system(GMENS) was employed to evaluate brain activity by using blood oxygen level dependent (BOLD) contrast. Gradient Echo EPI sequence with TR/TE= 2250/40 msec was used for 17 contiguous axial slices of 7mm thickness, covering the whole brain volume (240mm Field of view, 64 ${\times}$ 64 in plane resolution). The acquired data were applied to SPM99 for the processing such as realignment, normalization, smoothing, statistical ANOVA and statistical preference. Results/Disscusion: The comparison of familiar faces vs unfamiliar faces yielded significant activations in the medial temporal regions, the occipito temporal regions and in frontal regions. These results suggest that when volunteers are asked to recognize familiar faces among unfamiliar faces they tend to activate several regions frequently involved in face perception. The medial temporal regions are also activated for familiar and unfamiliar faces. This interesting result suggests a contribution of this structure in the attempt to match perceived faces with pre existing semantic representations stored in long term memory.

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