• 제목/요약/키워드: Diffusion in DNA

검색결과 75건 처리시간 0.023초

순무 모자이크 바이러스(TuMV)의 새로운 기주식물 탐색 (New Host Plants of Turnip Mosaic Potyvirus in Korea)

  • 최준근;윤주연;이세원;최장경
    • 한국식물병리학회지
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    • 제14권6호
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    • pp.625-629
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    • 1998
  • Turnip mosaic potyviruses (TuMV) were isolated from Rorippa indica and Armoracia lapathifolia showing mosaic symptoms in field. Identification of the TuMVs were carried out by host reactions of indicator plants, electron micrograph, serological properties and reverse transcription-poly-merase chain reaction (RT-PCR). Both viruses systemically infected Chenopodium quinoa, Nicotiana clevelandii, Brassica rapa, B. campestris subsp. pekinensis, B. juncea and Raphanus sativus, and developed local infection on inoculated leaves of C. quinoa, C. amaranticola, C. album, N. tabacum cv. Xanthi nc and Gomphrena grobosa. However, the viruses did not infect on N. glutinosa, Cucumis sativus and Vigna unguiculata. The filamentous particles, about 720 nm in length, and inclusion bodies were observed from the infected leaf tissues by dipping on electron microscopy. Crude sap of leaf infected with the viruses was reacted positively with an antiserum of TuMV in agar gel double diffusion. For detection of the viruses, RT-PCR was carried out with TuMV--specfic oligonucleotide primer. The RT-PCR products, a 1,092 bp DNA fragment, were obtained from naturally infected leaves of R. indica and A. lapathifolia. In inoculation test to seven cruciferous weeds with TuMV, infection occurred in Arabis glabra, Barbarea orthoceras, Capsella bursa-pastoris, Draba nomorosa var. hebecarpa, Rorippa cantoniensis and Thlaspi arvense.

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Antibacterial Activity of Recombinant Pig Intestinal Parasite Cecropin P4 Peptide Secreted from Pichia pastoris

  • Song, Ki-Duk;Lee, Woon-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권2호
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    • pp.278-283
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    • 2014
  • Cecropins (Cec) are antibacterial peptides and their expression is induced in a pig intestinal parasite Ascaris suum by bacterial infection. To explore the usefulness of its activity as an antibiotic, CecP4 cDNA was prepared and cloned into the pPICZ B expression vector and followed by the integration into AOX1 locus in Pichia pastoris. The supernatants from cell culture were collected after methanol induction and concentrated for the test of antimicrobial activity. The recombinant P. patoris having CecP4 showed antimicrobial activity when tested against Staphyllococcus aureus in disc diffusion assay. We selected one of the CecP4 clones (CecP4-2) and performed further studies with it. The growth of recombinant P. pastoris was optimized using various concentration of methanol, and it was found that 2% methanol in the culture induced more antibacterial activity, compared to 1% methanol. We extended the test of antimicrobial activity by applying the concentrated supernatant of CecP4 culture to Pseudomonas aeruginosa and E. coli respectively. Recombinant CecP4 also showed antimicrobial activity against both Pseudomona and E. coli, suggesting the broad spectrum of its antimicrobial activity. After improvements for the scale-up, it will be feasible to use recombinant CecP4 for supplementation to the feed to control microbial infections in young animals, such as piglets.

해양유출기름의 확산 시뮬레이션 모델 개발I- 폐쇄만에서의 3차원 흐름특성분석 - (Development of Simulation Model for Diffusion of Oil Spill in the Ocean 1 -Three Dimensional Characteristics of the Circulation in the Nearly Closed Bay-)

  • 이중우;김기철;강신영;도덕희
    • 한국항만학회지
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    • 제11권2호
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    • pp.241-255
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    • 1997
  • 한국의 부산에 위치한 감천만에서의 해수유동을 파악하기 위하여 3차원 유동모델을 사용하여 계산을 하고 그 결과를 관측치와 비교하였다. 모델은 개방경계에서의 조직, 바람, 화력발전소에서의 온배수에 의해 힘을 받아 해수유동을 재현한다. 난류 혼합계수는 $\kappa-\varepsilon$난류클로즈 모델의 의해 계산하였다. 수온, 염분, 해조류를 현장관측하여 관측자료와 모델결과를 비교하였다. 소용돌이 형태의 흐름이 현장관측자료와 모델결과에서 관측되었는데 이는 약한 조류와 풍성류, 온배수의 상호작용에 의해 형성된 것으로 분석되어진다. 표층과 저층의 흐름은 서로 연관되어 있는데 즉, 표층의 흐름이 강해지는 저츠으이 흐름이 약해지고 반대의 경우도 성립한다.

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Identification of Culturable Bioaerosols Collected over Dryland in Northwest China: Observation using a Tethered Balloon

  • Chen, Bin;Kobayashi, Fumihisa;Yamada, Maromu;Kim, Yang-Hoon;Iwasaka, Yasunobu;Shi, Guang-Yu
    • Asian Journal of Atmospheric Environment
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    • 제5권3호
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    • pp.172-180
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    • 2011
  • The transfer of microorganisms is important process for ecosystems. Microorganisms in dryland can transport itself to wetland through atmospheric diffusion, but only few papers reported about the atmospheric bioaerosol present over dryland. We carried out the direct sampling using a tethered balloon over Dunhuang City, China's northwestern dryland. Bioaerosols were collected using a tethered balloon with a bioaerosol collector at 820 m above the ground (1,960 m above the sea level) around noon on August 17, 2007. The bioaerosols were cultured after the collection at Dunhuang Meteorological observatory. Two strains of molds were isolated using the Nutrient agar medium. About 400-bp 18S rRNA partial sequences were amplified by PCR and determined afterwards. The results of a homology search by 18S rRNA sequences of isolates in DNA databases (GenBank, DDBJ, and EMBL) and an observation of the form revealed that two bioaerosols in the convective mixed layer over Dunhuang City were Cladosporium sp. and Aspergillus sp.

Biosurfactant Production by Marine Actinomycetes Isolates Streptomyces althioticus RG3 and Streptomyces californicus RG8 as Promising Sources of Antimicrobial and Antifouling Effects

  • Hamed, Moaz M.;Abdrabo, Mohamed A.A.;Youssif, Asmaa M.
    • 한국미생물·생명공학회지
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    • 제49권3호
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    • pp.356-366
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    • 2021
  • Two marine actinobacterial isolates, RG3 and RG8, were identified using 16Sr DNA as Streptomyces althioticus RG3 and Streptomyces californicus RG8 and submitted to the database of genetic information with accession numbers MW661230 and MW661234, respectively; they were found to have emulsification indexes of 60 ± 2.5% and 53 ± 2.2%, respectively. The biosurfactants obtained were stable at a temperature of 35℃ for both strains; they were stable at 10% NaCl, in the case of S. althioticus RG3 and at 10-15% NaCl in the case of Str.californicus RG8; both strains produced the most biosurfactant when exposed to alkaline conditions. We characterized the biosurfactants, including features such as their chemical composition, using Fourier transform infrared spectroscopy analysis. The antimicrobial activity of the biosurfactant extracts was evaluated using the well diffusion method against Vibrio alginolyticus MK170250, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 4027, and Staphylococcus aureus ATCC 25923. S. althioticus RG3 biosurfactants were found to have better antimicrobial activity than those of Str. californicus RG8, indicating that they may be used in pharmaceutical industries and in the manufacture of antifouling products.

육류용 고기로부터 분자진단을 이용한 항생제내성 유전자 양상 (Molecular detection of blaVIM, blaBIC, blaKPC, and blaSIM genes from isolated bacteria in retail meats)

  • 황유진
    • 한국산학기술학회논문지
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    • 제22권6호
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    • pp.413-419
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    • 2021
  • 본 연구의 목적은 확장 스펙트럼 𝛽-락타마제(ESBL) 항생제가 그람 음성 세균에 의한 감염을 치료하고 예방하는데 사용하고 있으며 임상에서 심각한 감염을 치료하는데 선택하는 마지막 옵션 역할을 한다. 미생물의 확장 스펙트럼 𝛽-락타마제(ESBL) 및/또는 카르바페네마제(Carbapenemase) 내성에 대한 보고는 전 세계적으로 확산되는 것으로 보고되며 항생제 치료에 많은 제한을 주는 요인으로 다약재 내성과 관련이 있기 때문에 보건 서비스에 큰 관심을 가지고 있다. 본 연구는 국내 시장에서 구매하여 분리한 육류로부터 세균을 분리 동정하여 항생제 저항성 테스트인 디스크 확산법을 사용하여 내성균을 분리 실험하였고, PCR과 DNA 시퀜싱방법을 수행아였다. 결과는 PCR을 수행하여 항생제 내성유전자와 유전자를 생산하는 ESBL의 존재를 검출하고 결과를 얻었다. 총 36개의 샘플 육류로부터 181개의 각각 분리된 세균을 추출하여 실험결과을 얻었다. 결과는 PCR과 DNA 염기서열을 분석하여 항생제내성 유전자로 blaVIM, blaBIC, blaKPC, blaSIM으로 나타났다. 분리한 육류 속의 박테리아는 별도 유전자 서열분석으로 4개의 다른 박테리아가 확인되었다. 이러한 결과는 소매되는 육류에서 발견되는 박테리아에 ESBL 내성유전자인 blaVIM, blaBIC, blaKPC, blaSIM를 가진 박테리아 균주가 있을 수 있으며 이는 특수 확장 스펙트럼 𝛽-락타마제(ESBL) 및/또는 카르바페네마제(Carbapenemase) 내성유전자가 확산될 수 있다는 것을 시사한다.

토끼풀에서 분리한 Peanut stunt virus의 성질 (Some Properties of an Isolate of Peanut stunt virus Isolated from White Clover (Trifolium repens L.))

  • 정구호;전용운;최장경;홍진성;류기현;이상용
    • 식물병연구
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    • 제14권1호
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    • pp.71-75
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    • 2008
  • 전형적인 모자이크 증상을 나타낸 토끼풀로부터 Peanut stunt virus(PSV)를 분리하여 Tr-PSV로 명명하였다. 이 연구에서는 Tr-PSV의 특성을 기주반응, 혈청학적 성질, dsRNA분석, RT-PCR 및 RFLP 분석 등을 통하여 지금까지 잘 알려진 ER-PSV Fny-CMV 및 LS-CMV와 비교하였다. Tr-PSV는 접종한 Nicotiana속 식물에서는 모두 전신감염되었으며, C. amaranticolor에서는 접종엽에 국부병반을 형성하였다. 그러나 동부에서는 대조로 이용한 ER-PSV와 마찬가지로 전신감염되어, 접종엽에 국부병반을 형성하는 대조의 CMV계통과는 구분되었다. Tr-PSV에 감염된 N. benthamiana 잎으로부터 추출한 dsRNA는 대조의 PSV나 CMV와 유사한 분자 크기의 4종의 dsRNA가 확인되었고, satellite RNA의 존재는 인정되지 않았다. Cucumovirus 특이적 프라이머를 이용하여 증폭시킨 PCR산물은 약 950 bp의 cDNA가 얻어졌고, 이를 이용하여 조사한 RFLP패턴은 ER-PSV와 같았다. 이와 같은 RFLP분석과 Er-PSV의 항혈청을 이용한 혈청학적 성질의 결과로부터 Tr-PSV는 ER-PSV와 같은 서브그룹 I에 속하는 PSV의 한 계통으로 판단되었다. 국내에서 토끼풀로부터 PSV가 분리 동정된 것은 이 논문이 처음이다.

Design, Optimization and Validation of Genomic DNA Microarrays for Examining the Clostridium acetobutylicum Transcriptome

  • Alsaker, Keith V.;Paredes, Carlos J.;Papoutsakis, Eleftherios T.
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권5호
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    • pp.432-443
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    • 2005
  • Microarray technology has contributed Significantly to the understanding of bacterial genetics and transcriptional regulation. One neglected aspect of this technology has been optimization of microarray-generated signals and quality of generated information. Full genome microarrays were developed for Clostridium acetobutylicum through spotting of PCR products that were designed with minimal homology with all other genes within the genome. Using statistical analyses it is demonstrated that Signal quality is significantly improved by increasing the hybridization volume. possibly increasing the effective number of transcripts available to bind to a given spot, while changes in labeled probe amounts were found to be less sensitive to improving signal quality. In addition to Q-RT-PCR, array validation was tested by examining the transcriptional program of a mutant (M5) strain lacking the pSOL1 178-gene megaplasmid relative to the wildtype (WT) strain. Under optimal conditions, it is demonstrated that the fraction of false positive genes is 1% when considering differentially expressed genes and 7% when considering all genes with signal above background. To enhance genomic-scale understanding of organismal physiology, using data from these microarrays we estimated that $40{\sim}55%$ of the C. acetobutylicum genome is expressed at any time during batch culture, similar to estimates made for Bacillus subtilis.

양송이 배지로부터 분리된 Bacillus subtilis NO12의 특성 (Isolation and characterization of Bacillus subtilis NO12 from button mushroom substrates)

  • 김혜수;박현영;이찬중;공원식;조수정
    • 한국버섯학회지
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    • 제15권4호
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    • pp.249-253
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    • 2017
  • Cellulase와 xylanase 분비능이 우수한 세균을 분리하기 위하여 부여군 석성면 지역의 양송이 재배농장으로부터 수확후배지를 수집하였다. 양송이 수확후배지로부터 12종의 균주를 분리하였으며 이 중 cellulase와 xylanase 활성이 가장 우수한 균주 NO12를 최종 선발하였다. 분리균 NO12의 생리적 생화학적 특성은 Bacillus ID kit와 MicroLog system을 이용하여 조사하였으며 분리균 NO12는 Bacillus subtilis와 유사한 특징을 나타내었다. 분리균 NO12의 16S rDNA 염기서열도 B. subtilis와 99.2%의 상동성을 나타내었다. 이와 같은 결과를 종합하여 분리균 NO12는 B. subtilis NO12로 동정되었다. 분리균이 분비하는 cellulase와 xylanase 활성은 분리균이 증식함에 따라 대수증식기 중반부터 급격히 증가하였고 정지기에 진입하면 효소활성이 더 이상 증가하지 않는 것으로 나타났으며 xylanase 활성은 대수증식기 초기부터 지속적으로 증가하여 대수증식기 중반에 최대활성을 나타내었다.

Isolation, Identification and Optimal Culture Conditions of Streptomyces albidoflavus C247 Producing Antifungal Agents against Rhizoctonia solani AG2-2

  • Islam, Rezuanul;Jeong, Yong-Tae;Ryu, Yeon-Ju;Song, Chi-Hyun;Lee, Yong-Se
    • Mycobiology
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    • 제37권2호
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    • pp.114-120
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    • 2009
  • Streptomyces albidoflavus C247 was isolated from the soil of the Gyeongsan golf course in Korea. Physiological, biochemical and 16S rDNA gene sequence analysis strongly suggested that the isolate belonged to Streptomyces albidoflavus. Preliminary screening revealed that the isolate was active against fungi and bacteria. Self-directing optimization was employed to determine the best combination of parameters such as carbon and nitrogen source, pH and temperature. Nutritional and culture conditions for the production of antibiotics by this organism under shake-flask conditions were also optimized. Maltose (5%) and soytone (5%) were found to be the best carbon and nitrogen sources for the production of antibiotics by S. albidoflavus C247. Additionally, 62.89% mycelial growth inhibition was achieved when the organism was cultured at $30^{\circ}C$ and pH 6.5. Ethyl acetate (EtOAc) was the best extraction solvent for the isolation of the antibiotics, and 100 ${mu}$/ml of EtOAc extract was found to inhibit 60.27% of the mycelial growth of Rhizoctonia solani AG2-2(IV) when the poison plate diffusion method was conducted.