• Title/Summary/Keyword: merA

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An Oligonucleotide Microarray Bait for Isolation of Target Gene Fragments

  • Shi, Rong;Ma, Wen-li;Liu, Cui-Hua;Song, Yan-Bin;Mao, Xiang-Ming;Zheng, Wen-Ling
    • BMB Reports
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    • v.37 no.2
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    • pp.148-152
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    • 2004
  • A new molecular-baiting method was studied by retrieving targeted gene fragments from an oligonucleotide microarray bait after hybridization. To make the microarray bait, 70-mer oligonucleotides that were designed to specifically represent the SSA1 gene of Saccharomyces cerevisiae were printed on the slide. Samples of the Saccharomyces cerevisiae mRNA were extracted and labeled by the RD-PCR (Restriction Display PCR) method using the Cy5-labelled universal primer, then applied for hybridization. The sample fragments that hybridized to the microarray were stripped, and the eluted cDNAs were retrieved and cloned into the pMD 18-T vector for transformation, plasmid preparation, and sequencing. BLAST searching of the GenBank database identified the retrieved fragments as being identical to the SSA1 gene (from 2057-2541bp). A new method is being established that can retrieve the sample fragments using an oligo-microarray-bait.

Investigation of Experimental Results Using the Drying Model for a Heat Pump Dryer (열펌프 건조 해석 모델을 이용한 측정 결과의 분석)

  • Lee, Kong-Hoon;Kim, Ook-Joong;Kim, Jong-Ryul;Lee, Sang-Ryoul
    • Proceedings of the KSME Conference
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    • 2008.11b
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    • pp.2268-2273
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    • 2008
  • The drying model has been used to obtain the fundamental information required to design the heat pump dryer with the simple thermodynamic model. In the model, the input conditions are crucial to obtain the acceptable results. The model includes one-stage heat pump cycle, simple drying process using the drying efficiency. The drying efficiency is defined with the conditions of inlet and outlet in the dryer. The experiment has been carried out in the pilot dryer with one-stage heat pump cycle. Refrigerant 134a is used in the heat pump cycle. In the dryer, some of drying air flows through the heat pump system and the rest of air bypasses the heat pump system and circulates through the drying chamber. Some operating conditions from the pilot dryer are used as input conditions of the model and the results are compared with experimental results for the validation.

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Influence of the N- and C-Terminal Regions of Antimicrobial Peptide Pleurocidin on Antibacterial Activity

  • Cho, Jaeyong;Choi, Hyemin;Lee, Dong Gun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1367-1374
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    • 2012
  • Pleurocidin, a 25-mer antimicrobial peptide, has been known to exhibit potent antibacterial activity. To investigate the functional roles in N- and C-terminal regions of pleurocidin on the antibacterial activity, we designed four truncated analogs. The antibacterial susceptibility testing showed that pleurocidin and its analogs exerted antibacterial effect against various bacterial strains and further possessed specific activity patterns corresponding with their hydrophobic scale [pleurocidin > Anal 3 (1-22) > Anal 1 (4-25) > Anal 4 (1-19) > Anal 2 (7-25)]. Fluorescence experiments using 1,6-diphenyl-1,3,5-hexatriene (DPH) and 3,3'-dipropylthiadicarbocyanine iodide [$diSC_3(5)$] indicated that the differences in antibacterial activity of the peptides were caused by its membrane-active mechanisms including membrane disruption and depolarization. Blue shift in tryptophan fluorescence demonstrated that the decrease in net hydrophobicity attenuates the binding affinity of pleurocidin to interact with plasma membrane. Therefore, the present study suggests that hydrophobicity in the N- and C-terminal regions of pleurocidin plays a key role in its antibacterial activity.

A Fragmentation and Search Method of Query Document for Partially Plagiarized Section Detection (부분표절구간 검출을 위한 질의문서의 분할 및 탐색 기법)

  • Ock, Chang-Seok;Seo, Jong-Kyu;Cho, Hwan-Gue
    • Proceedings of the Korea Information Processing Society Conference
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    • 2012.11a
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    • pp.586-589
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    • 2012
  • 표절과 관련된 이슈가 주목받고 있는 상황에서 표절을 검출하는 방법에 대한 연구가 활발히 진행되고 있다. 일반적으로 표절구간 검출을 위해 복잡한 자연어처리와 같은 의미론적 접근방법이 아닌 비교적 단순한 어휘기반의 문자열 처리 방법을 사용한다. 대표적인 방법으로는 지문법 (Fingerprinting)과 서열정렬 (Sequence alignment) 등이 있다. 하지만 이 방법들을 이용하여 대용량 문서에 대한 표절검사를 수행하기에는 시공간적 복잡도의 문제가 발생한다. 본 논문에서는 이러한 단점을 극복하기 위해 NGS (Next Generation Sequencing)에서 사용하는 BWT (Burrows-Wheeler Transform)[1]를 이용한 탐색방법을 응용한다. 또한 부분표절구간을 검출하고 정확도를 향상시키기 위해 질의문서를 분할하여 작은 조각으로 만든 뒤, 조각들에 대한 질의탐색을 수행한다. 본 논문에서는 질의문서를 분할하는 두 가지 방법을 소개한다. 두 가지 방법은 k-mer analysis를 이용한 방법과 random-split analysis를 이용한 방법으로, 각 방법의 장단점을 실험을 통해 분석하고 실제 부분표절구간의 검출 정확도를 측정하였다.

Genetic Diversity Evaluation of Thamnocalamus spathiflorus (Trin.) Munro Accessions through Morphological and Randomly Amplified Polymorphic DNA (RAPD) Markers

  • Tiwari, Chandrakant;Bakshi, Meena;Gupta, Dinesh
    • Journal of Forest and Environmental Science
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    • v.35 no.2
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    • pp.90-101
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    • 2019
  • Biodiversity refers to the total number and variation among species of flora and fauna of an area. Due to tremendous biotic especially anthropogenic pressure these natural resources are being vanishing. In present study genetic diversity among accessions of Thamnocalamus spathiflorus was evaluated. A total of 51 vegetative characters and 42 primers (10-mer) were screened. Out of 42 screened primers, 28 polymorphic primers were selected for further analysis. A total of 263 bands were recorded as polymorphic whereas 48 bands were monomorphic. The resolving power (Rp) of 28 Randomly Amplified Polymorphic DNA (RAPD) primers ranged from 4.6 (OPE08) to 17.6 (OPA11). The polymorphic information content (PIC) value ranged from 0.21 (OPAH09) to 0.44 (OPG02). The result revealed high degree of genetic relatedness (56 to 80%). Cluster analysis revealed two major clusters both for morphology as well as RAPD. Unlike morphological characterization, the accession (D5) from Bahli, Rampur, Shimla (H.P.) was clustered separately from the others in RAPD cluster analysis. Accessions with closed locality grouped together through RAPD marker system however analogy was recorded for morphological traits. The study conducted reflects the utility of RAPD technique for species identification and phylogenetic studies in bamboo for conducting bamboo breeding program.

Molecular discrimination of Panax ginseng species

  • Um, Jae-Young;Chung, Hwan-Suck;Kim, Hyun-Ju;Kim, Dae-Ki;Shim, Kyung-Shik;Lee, Kang-Yong;Kim, Jeong-Sook;Choi, Tae-Jin;Kim, Nam-Song;An, Nyeon-Hyoung;Lee, Kang-Min;Lee, Young-Mi;Kim, Jeong-Joong
    • Advances in Traditional Medicine
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    • v.1 no.2
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    • pp.52-58
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    • 2000
  • In order to develop convenient and reproducible methods for identification of ginseng drugs at a DNA level, RAPD (randomly amplified polymorphic DNA) and PCR-RFLP (PCR-Restriction fragment length polymorphism) analysis were applied within Panax species. To authenticate Panax ginseng betvyeen Chinese and Korean ginseng population, RAPD analysis were carried out using 20 mer-random primer. The similarity coefficients among the DNA of ginseng plants analyzed were low, ranging from 0.197 to 0.491. In addition, using PCR-RFLP analysis, very different fingerprints were obtained within Korean ginseng plants. These results suggest that these methods are able to authenticate the concerned Panax species. Broader application of this approach to authenticate other morphologically similar medicinal materials is rationalized.

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Identification of mono- or poly-specific monoclonal antibody to Porphyromonas gingivalis heat-shock protein 60

  • Choi, Jeom-Il;Lee, Sang-Yull;Kim, Koan-Hoi;Choi, Bong-Kyu;Kim, Myung-Jin
    • Journal of Periodontal and Implant Science
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    • v.41 no.2
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    • pp.54-59
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    • 2011
  • Purpose: The aim of this study was to define the immunoreactive specificity of Porphyromonas gingivalis (P. gingivalis) heat shock protein (HSP) 60 in periodontitis and atherosclerosis. Methods: In an attempt to define the cross-reactive bacterial heat-shock protein with human self-antigen at molecular level, we have introduced a novel strategy for cloning hybridoma producing anti-P. gingivalis HSP 60 which is polyreactive to bacterial HSPs or to the human homolog. Results: Five cross-reactive clones were obtained which recognized the #19 peptide (TLVVNRLRGSLKICAVKAPG) among 37 synthetic peptides (20-mer, 5 amino acids overlapping) spanning the whole molecule of P. gingivalis HSP 60. We have also established three anti-P. gingivalis HSP 60 monoclonal antibodies demonstrating mono-specificity. These clones recognized the #29 peptide (TVPGGGTTYIRAIAALEGLK). Conclusions: Peptide #19 and #29 of P. gingivalis HSP 60 might be important immunoreactive epitopes in the immuno-pathogenic mechanism of bacterial antigen-triggered autoimmune diseases.

Mercury Resistance and Removal Mechanisms of Pseudomonas sp. Isolated Mercury-contaminated Site in Taiwan

  • Luo, Kai-Hong;Chen, Ssu-Ching;Liao, Hung-Yu
    • Journal of Soil and Groundwater Environment
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    • v.21 no.5
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    • pp.16-24
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    • 2016
  • A new strain of Pseudomonas sp. was isolated from mercury (Hg)-contaminated sites in Taiwan. This bacterium removed more than 80% of Hg present in the culture medium at 12 h incubation and was chosen for further analysis of the molecular mechanisms of Hg tolerance/removal abilities in this Pseudomonas sp. We used RNA-seq, one of the next-generation sequencing methods, to investigate the transcriptomic responses of the Pseudomonas sp. exposed to 60 mg/L of Hg2+. We de novo assembled 4,963 contigs, of which 10,533 up-regulated genes and 5,451 down-regulated genes were found to be regulated by Hg. The 40 genes most altered in expression levels were associated with tolerance to Hg stress and metabolism. Functional analysis showed that some Hg-tolerant genes were related to the mer operon, sulfate uptake and assimilation, the enzymatic antioxidant system, the HSP gene family, chaperones, and metal transporters. The transcriptome were analyzed further with Gene Ontology (GO) and Cluster of Orthologous Groups (COGs) of proteins and showed diverse biological functions and metabolic pathways under Hg stress.

The Influence of the N-Terminal Region of Antimicrobial Peptide Pleurocidin on Fungal Apoptosis

  • Choi, Hyemin;Lee, Dong Gun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1386-1394
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    • 2013
  • In our previous study, the 25-mer antimicrobial peptide pleurocidin (Ple) had been thought to induce apoptosis in Candida albicans. This study demonstrated that reactive oxygen species (ROS) production was a major cause of Ple-induced apoptosis. Four truncated analogs were synthesized to understand the functional roles in the N- and C-terminal regions of Ple on the apoptosis. Ple, Ple (4-25), Ple (1-22), and Ple (1-19) produced ROS, including hydroxyl radicals, on the order of [Ple > Ple (1-22) > Ple (4-25) > Ple (1-19)], whereas Ple (7-25) did not induce any ROS production. The results suggested that the N-terminal deletion affected the ROS-inducing activities much more than that of the C-terminal deletion, and net hydrophobicity [Ple > Ple (1-22) > Ple (4-25) > Ple (1-19) > Ple (7-25)] was related to ROS generation rather than other primary factors like net charge. Hence, we focused on the N-terminal-truncated peptides, Ple (4-25) and Ple (7-25), and examined other apoptotic features, including mitochondrial membrane depolarization, caspase activation, phosphatidylserine externalization, and DNA and nuclear fragmentation. The results also confirmed the disappearance of apoptotic activity of Ple (7-25) by the truncation of the N-terminal region (1-6) and the specific activity patterns between Ple and analogs. In conclusion, the N-terminal region of Ple played an important role in apoptosis.

Comparison of RAPD Profiles and Phenotypical Characters of Streptococcal Strains (연쇄상구균의 표현형적 특성과 RAPD profiles 비교)

  • Song, Jin-Gyeong;Kim, Jong-Hun;Kim, Eun-Hui
    • Journal of fish pathology
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    • v.16 no.1
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    • pp.51-59
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    • 2003
  • Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.