• Weed & Turfgrass Science
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• 제6권2호
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• pp.157-164
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• 2017

본 연구는 미생물을 이용하여 친환경적인 잔디관리를 위해 계분이나 돈분의 퇴비화 과정에서 얻어진 가축분 퇴비로부터 단백질 및 탄수화물 분해능과 잔디 갈색퍼짐병(large patch), 갈색마름병(brown patch), 그리고 동전마름병(dollar spot) 병원균에 항균활성을 보이는 미생물을 분리하기 위하여 실시하였다. 분리된 미생물은 총 68균주였고, 이들을 대상으로 단백질 분해활성, 탄수화물 분해활성 및 잔디 주요병원균에 대한 항균활성을 조사하여 활성이 높은 미생물 34균주를 선발하였다. 이 중에서 단백질과 탄수화물 분해 및 항균활성을 나타내는 균주인 ASC-14, ASC-18 및 ASC-35를 선발하였다. 이들 선발 균주를 대상으로 16s rRNA 유전자 분석 결과 ASC-14와 ASC-18은 B. amyloliquefaciens로 확인되었고, 반면에 ASC-35는 B. subtilis 세균으로 최종 동정되었다.

• 인터넷정보학회논문지
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• 제18권2호
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• pp.43-51
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• 2017

게임 산업이 발전하면서 콘텐츠의 생성 속도보다 훨씬 빠른 속도로 콘텐츠가 소비되고 있고, 플레이어의 게임 숙련도에 적합한 레벨의 게임 콘텐츠들이 지속적으로 제공될 것을 필요로 하고 있다. 이러한 문제를 효과적으로 해결하기 위해 활용되는 방법이 인공지능(Artificial Intelligence, AI)을 이용한 절차적 콘텐츠 생성(Procedural Content Generation, PCG)이다. 본 논문에서는 유전 알고리즘을 이용하여 플레이어에게 적합한 난이도를 가지고 있는 다양한 종류의 몬스터를 자동 생성하는 절차적 방법을 제안한다. 몬스터들의 주요 속성을 유전자로 구성하고 다양한 종류의 몬스터 유전자들로 염색체를 만들어 이용한다. 생성된 몬스터와 플레이어의 전투 시뮬레이션으로 유전자를 평가하여 선택 후 교배한다. 본 논문의 제안 방법을 이용해 플레이어 적응형 몬스터들을 유전 알고리즘에 기반을 두어 절차적으로 생성하고, 염색체 개수에 따라 생성된 몬스터의 다양성을 비교해본다.

• The Plant Pathology Journal
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• 제31권1호
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• pp.33-40
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• 2015

Pigeonpea Sterility Mosaic Disease (PSMD) is an important foliar disease caused by Pigeonpea sterility mosaic virus (PPSMV) which is transmitted by eriophyid mites (Aceria cajani Channabasavanna). In present study, a F2 mapping population comprising 325 individuals was developed by crossing PSMD susceptible genotype (Gullyal white) and PSMD resistant genotype (BSMR 736). We identified a set of 32 out of 300 short decamer random DNA markers that showed polymorphism between Gullyal white and BSMR 736 parents. Among them, eleven DNA markers showed polymorphism including coupling and repulsion phase type of polymorphism across the parents. Bulked Segregant Analysis (BSA), revealed that the DNA marker, IABTPPN7, produced a single coupling phase marker (IABTPPN $7_{414}$) and a repulsion phase marker (IABTPPN $7_{983}$) co-segregating with PSMD reaction. Screening of 325 F2 population using IABTPPN7 revealed that the repulsion phase marker, IABTPPN $7_{983}$, was co-segregating with the PSMD responsive SV1 at a distance of 23.9 cM for Bidar PPSMV isolate. On the other hand, the coupling phase marker IABTPPN $7_{414}$ did not show any linkage with PSMD resistance. Additionally, single marker analysis both IABTPPN $7_{983}$ (P<0.0001) and IABTPPN $7_{414}$ (P<0.0001) recorded a significant association with the PSMD resistance and explained a phenotypic variance of 31 and 36% respectively in $F_2$ population. The repulsion phase marker, IABTPPN7983, could be of use in Marker-Assisted Selection (MAS) in the PPSMV resistance breeding programmes of pigeonpea.

• Journal of Plant Biotechnology
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• 제30권4호
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• pp.307-313
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• 2003

파리지옥풀 first trap leaf에만 발현하는 유전자군을 탐색하기 위하여 기내배양 식물체와 포충능력을 가진 3년생 실생주을 이용하여 각각의 포충잎 (leaf base), 꽃조직 (flower tissue) 및 포충잎트랩 (first trap leaf)으로부터 분리한 RNA로 Fluorescent differential display (FDD)를 실시하였다. First trap leaf특이발현 유전자 15개를 screening하여 염기서열을 분석하였다. 분리된 DNA들은 protease inhibitor (Pl), myo-inositol-1-phosphate synthase 및 lipocalin-type prostaglandin D syn-thase 유전자들과 매우 유사하였다. 또한 Northern blot분석 결과, 이들 유전자들이 first trap leaf에 특이적으로 발현하고 있는 것을 확인하였다 FDD방법은 세포, 조직 및 기관에 특이적으로 발현하고 있는 유전자들을 선발하는데 매우 유용한 수단으로 사용될 수 있다.

• Journal of Microbiology
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• 제39권4호
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• pp.300-304
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• 2001

Epstein-Barr Virus(EBV)-transformed lymphoblastoid B cell lines, BLCL which expresse antigens, are potential antigen-presenting cells(APCs) for the induction of CTL in vitro. However transfection of BLCLs with subsequent selection by antibiotics is notoriously difficult because plating efficiencies of BLCLsare reported to be 1% or less. To generated stable transfectants of BLCLs we produced high titers of retroviruess encoding pp 65 antigen of human cytomegalovirus of foreign antigens and trans-duced them of BLCLs. The pp 65 gene was cloned into the retroviral vector pLXSN. The recombinant retroviral vector was transfected to ecotropic packaging cell line, CP&E86, and this polyclonal recom-binant retrovirus was transduced to PA317 that is amphotropic pakaging cell line. The titers of colned PA317 amphotropic retroviruses ranged from 5 to $\times$10$^{6}$ colony forming units (CFU)per ml (CFU/ml) We performed three rounds of consecutive transductions to BLCLs in order to improve the clon-ing effieiencies. The expression of recombinant HCMV-pp65 antigen was more than 20% after the final transduction. THe third-transduced BLCLs were easily selected in optimal concentration of G418. BLCLs expressing foreign antigens could be used as target cells for CTL assay and/or as APCs for induction of in vitro CTL responses specific for viral and tumor antigens.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1100-1108
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• 2009

Lactic acid bacteria (LAB) isolated from various fruits and vegetables were screened in order to determine appropriate fermentation starters for manufacturing functional fermented onion juice. From the initial screening test comprising more than 700 isolated LAB, 16 isolates were selected based on their acid production rate. Among the selected isolates, the fermentation broth of KC-007 exhibited the highest electron donating and nitrite scavenging activities, with values at pH 1.2 of 95.6 and 68.7%, respectively. From the overall results obtained in this study, we finally selected the bacterium KC-007 as a fermentation starter. This bacterium was identified and named as Pediococcus pentosaceus based on its morphological and physiological characteristics, carbon-utilization pattern (as assessed using an API 50CHL kit), and molecular genetic characteristics (as assessed using the nucleotide sequence of the 16S rRNA gene). The optimal temperature, pH, and starter inoculation concentration (v/v) required for growth of the isolated strain were $40^{\circ}C$, pH 4.0-6.0, and 2%(v/v), respectively.

• 한국미생물·생명공학회지
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• 제20권3호
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• pp.263-270
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• 1992

채소 등 작물의 부패균인 Erwinia carotovora subsp. carotovora의 생육을 억제하는 길항세균을 경작지 표토층으로부터 분리하고 배양조건에 따른 억제력변화를 조사하고, 길항관련 유전자의 소재를 확인하였다. 분리균들중 가장 우수한 억제력을 갖는 S4, S14, S65 균주를 최종 선발하고 Pseudomonas 근연종으로 동정하였다. 523 배지를 사용하여 배양조건에 따른 억제력 변화를 조사한 결과 배지초기 pH를 7-8로 조정하여 $30^{\circ}C$에서 3일간 배양하였을 때 가장 높은 억제력을 보였고, 배지성분중 탄소원으로 mannitol과 sorbitol, 무기염으로 CaCl2를 첨가했을 때 효과적이었으나 질소원에 대한 감수성은 낮았다.

• 한국미생물·생명공학회지
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• 제35권2호
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• pp.98-103
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• 2007

산란계의 분변으로부터 생균제 특성을 가진 유산균을 분리하기 위하여, 무작위 선발법과 agar well diffusion assay법을 사용하여 다수의 유산균을 1차 선발하였으며 이 중에서 대장균 억제능이 가장 우수한 CPM-7 균주를 분리하였다. 최종 선발된 CPM-7 균주는 형태학적 특징, 당 이용성 및 16S rRNA 서별 분석을 통하여 Lactobacillus salivarius CPM-7으로 동정되었다. L. salivarius CPM-7은 내산성 및 내담즙성이 우수한 것으로 나타났는데, pH 2에서 30분 동안 및 pH 3에서 6시간 동안 생균수가 거의 유지되었으며, bile salt 0.2%가 첨가된 MRS 배지에서 증식할 수 있었다. L. salivarius CPM-7은 a-galactosidase를 포함한 다수의 효소를 생산하는 것으로 확인되었으며, 돼지의 장 상피세포에 흡착할 수 있었다. L. salivarius CPM-7의 배양액 및 중화액은 자돈 설사를 유발하는 것으로 알려진 E. coli K88에 대한 강력한 억제능을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.511-520
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• 2015

Triclosan, the widely used biocide, specifically targets enoyl-acyl carrier protein reductase (ENR) in the bacterial fatty acid synthesis system. Although the fish pathogen Aeromonas salmonicida subsp. salmonicida exhibits triclosan resistance, the nature of this resistance has not been elucidated. Here, we aimed to characterize the triclosan resistance of A. salmonicida subsp. salmonicida causing furunculosis. The fosmid library of triclosan-resistant A. salmonicida subsp. salmonicida was constructed to select a fosmid clone showing triclosan resistance. With the fosmid clone showing triclosan resistance, a subsequent secondary library search resulted in the selection of subclone pTSR-1. DNA sequence analysis of pTSR-1 revealed the presence of a chromosomal-borne fabV-encoding ENR homolog. The ENR of A. salmonicida (FabVas) exhibited significant homology with previously known FabV, including the catalytic domain YX₍₈₎K. fabVas introduction into E. coli dramatically increased its resistance to triclosan. Heterologous expression of FabVas might functionally replace the triclosan-sensitive FabI in vivo to confer E. coli with triclosan resistance. A genome-wide search for fabVas homologs revealed the presence of an additional fabV gene (fabVas2) paralog in A. salmonicida strains and the fabVas orthologs from other gram-negative fish pathogens. Both of the potential FabV ENRs expressed similarly with or without triclosan supplement. This is the first report about the presence of two potential FabV ENRs in a single pathogenic bacterium. Our result suggests that triclosan-resistant ENRs are widely distributed in various bacteria in nature, and the wide use of this biocide can spread these triclosan-tolerant ENRs among fish pathogens and other pathogenic bacteria.

• Journal of Microbiology and Biotechnology
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• 제20권10호
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• pp.1463-1470
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• 2010

E. coli has long been widely used as a host system for the manufacture of recombinant proteins intended for human therapeutic use. When considering the impurities to be eliminated during the downstream process, residual host cell DNA is a major safety concern. The presence of residual E. coli host cell DNA in the final products is typically determined using a conventional slot blot hybridization assay or total DNA Threshold assay. However, both the former and latter methods are time consuming, expensive, and relatively insensitive. This study thus attempted to develop a more sensitive real-time PCR assay for the specific detection of residual E. coli DNA. This novel method was then compared with the slot blot hybridization assay and total DNA Threshold assay in order to determine its effectiveness and overall capabilities. The novel approach involved the selection of a specific primer pair for amplification of the E. coli 16S rRNA gene in an effort to improve sensitivity, whereas the E. coli host cell DNA quantification took place through the use of SYBR Green I. The detection limit of the real-time PCR assay, under these optimized conditions, was calculated to be 0.042 pg genomic DNA, which was much higher than those of both the slot blot hybridization assay and total DNA Threshold assay, where the detection limits were 2.42 and 3.73 pg genomic DNA, respectively. Hence, the real-time PCR assay can be said to be more reproducible, more accurate, and more precise than either the slot blot hybridization assay or total DNA Threshold assay. The real-time PCR assay may thus be a promising new tool for the quantitative detection and clearance validation of residual E. coli host cell DNA during the manufacturingprocess for recombinant therapeutics.