• 제목/요약/키워드: Selective isolation

검색결과 171건 처리시간 0.026초

Development of Molecular Biological Methods to Analyze Bacterial Species Diversity in Freshwater and Soil Ecosystems

  • Lee, Dong-Hun;Noh, Sung-Ae;Kim, Chi-Kyung
    • Journal of Microbiology
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    • 제38권1호
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    • pp.11-17
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    • 2000
  • A new method was developed for the rapid analysis of diverse bacterial species in the natural environment. Our method is based on PCR-single-strands-conformation polymorphism (PCR-SSCP) and selective isolation technique of single-stranded DNA. Variable V3 fragments of 16S rDNA were amplified by PCR with bacterial 16S rDNA primers, where one of the primers was biotinylated at the 5'-end. The biotinylated strands of the PCR products were selectively isolated by using streptavidin paramagnetic particles and a magnetic stand, to prevent SSCP analysis producing heteroduplexes from heterogeneous DNA samples. The selected strands were separated by electrophoresis on a polyacrylamide gel, and detected by silver staining. Analysis of PCR products from 8 bacterial strains demonstrated their characteristic DNA band patterns. In addition, changes in the structure of the bacterial community and species diversity in the microcosm treated with phenol could be monitored. After 3 weeks of incubation, phenol and its intermediate, 2-hydroxy-muconic-semialdehyde, were degraded by indigenous bacteria. These dominating bacterial populations were identified as strong bands on an SSCP gel. Therefore, this study provides useful tools for microbial community analysis of natural habitats.

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STI-CMP 공정 적용을 위한 연마 정지점 고찰 (A Study of End Point Detection Measurement for STI-CMP Applications)

  • 김상용;서용진
    • 한국전기전자재료학회논문지
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    • 제14권3호
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    • pp.175-184
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    • 2001
  • In this study, the improved throughput and stability in device fabrication could be obtained by applying CMP process to STi structue in 0.18 um semiconductor device. To employ the CMP process in STI structure, the Reverse Moat Process used to be added after STI Fill, as a result, the process became more complex and the defect were seriously increased than they had been,. Removal rate of each thin film in STI CMP was not uniform, so, the device must have been affected. That is, in case of excessive CMP, the damage on the active area was occurred, and in the case of insufficient CMP nitride remaining was happened on that area. Both of them deteriorated device characteristics. As a solution to these problems, the development of slurry having high removal rate and high oxide to nitride selectivity has been studied. The process using this slurry afford low defect levels, improved yield, and a simplified process flow. In this study, we evaluated the 'High Selectivity Slurry' to do a global planarization without reverse moat step, and also we evaluated EPD(Eend Point Detection) system with which 'in-situ end point detection' is possible.

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Evaluation of ${\mu}$-Calpain Inhibitory Activity of Korean Indigenous Marine Organism Extracts

  • Lee, Yoo-Jin;Lee, Eun-Young;Han, Ah-Reum;Song, Jun-Im;Kwon, Young-Joo;Seo, Eun-Kyoung
    • Natural Product Sciences
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    • 제18권2호
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    • pp.102-105
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    • 2012
  • Marine organism extracts were prepared from 26 species of Korean indigenous marine organisms, including 25 species belonging in class Anthozoa of phylum Cnidaria and a species belonging to subphylum Urochordata of phylum Chordata, and screened their inhibitory effects against ${\mu}$-calpain. As a result, the thirteen extracts were found to be active in the criteria of $IC_{50}$ < 100 ${\mu}g/ml$. Among them, the MeOH extracts of Plexauroides praelonga and Alveopora japonica showed remarkable ${\mu}$-calpain inhibitory activity with $IC_{50}$ values of $4.62{\pm}0.22$ and $4.82{\pm}0.07{\mu}g/ml$, respectively. In addition, chemical investigation of A. japonica led to the isolation of an active compound, hexadecyl tetradecanoate, as a selective cathepsin B inhibitor ($IC_{50}=9.05{\pm}2.45{\mu}M$). This compound was isolated as constituent of A. japonica for the first time in the present study.

Isolation of Sangivamycin from Streptomyces sp. A6497 and its Herbicidal Activity

  • HWANG EUI IL;YUN BONG SIK;CHOI SUNG WON;KIM JIN SEOG;LIM SE JIN;MOON JAE SUN;LEE SANG HAN;KIM SUNG UK
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.434-437
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    • 2005
  • During the screening for the inhibitors of cellulose biosynthesis as herbicides, we discovered a Streptomyces sp. A6497 with a selective antifungal activity against cellulose containing Phytophthora parasitica, but not against cellulose lacking Candida albicans. The inhibitor was isolated and identified, using a series of chromatographies. Based on structure analyses with UV spectrophotometry, mass and various NMR, the inhibitor was identified as sangivamycin. The compound exhibited strong antifungal activities against P. parasitica (MIC; 3.125 $\mu$g/ml). In particular, it showed strong herbicidal activities against various weeds in the greenhouse experiment. Taken together, these results suggest that sangivamycin is a useful lead compound for the development of new herbicides.

Design, Synthesis and Preliminary Biological Evaluation of a Biotin-S-S-Phosphine Reagent

  • Kang, Dong W.;Kim, Eun J.
    • Bulletin of the Korean Chemical Society
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    • 제35권2호
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    • pp.383-391
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    • 2014
  • Biotin-S-S-Phosphine was designed and synthesized as a potential tool for a proteomic study of O-GlcNAcmodified proteins. This reagent features a disulfide linker between a triarylphosphine moiety, which allows selective conjugation to azide-containing proteins, and a biotin moiety that can allow easy isolation through its strong affinity toward avidin-coated solid beads. The disulfide linkage within this reagent can allow the easy release of the bound molecules of interest, which is difficult to achieve when a biotin:avidin pair is used alone, by reducing the disulfide bond of the reagent with DTT. Preliminary in vitro biological assays with azidelabeled and unlabeled cell lysates and a pure protein Nup62 showed that the Biotin-S-S-Phosphine reagent is highly reactive toward the free thiol groups of proteins. When a molecular tool with a disulfide linker is applied to the enrichment of the molecules of interest from other species, it is important to block the free-thiols of the sample using exhaustive alkylation prior to the Staudinger ligation reactions to restore the bioorthogonal nature of this reaction.

Seed and Root Rots of Ginseng (Panax quinquefolius L) Caused by Cylindrocarpon destructans and Fusarium spp.

  • Reeleder, R.D.;Roy, R.;Capell, B.
    • Journal of Ginseng Research
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    • 제26권3호
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    • pp.151-158
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    • 2002
  • Ginseng (Panax quinquefolius L.) has become one of the most valuable herb crops grown in North America. However, traditional cropping practices are favourable to disease and significant losses due to root disease are common, despite frequent use of fungicides. Seedlots are often contaminated with pathogens, however, little is known about the causes of seed decay and the role of seed pathogens as incitants of root rots. It was shown that both Fusarium spp. and Cylindrocarpon destructans were able to rot seeds and that C. destructans was more virulent than Fusarium spp. on seedling roots. A modified rose bengal agar MRBA) medium (1 g KH$_2$PO$_4$; 0.5 g MgSO$_4$; 50 mg rose bengal; 10 g dextrose; 5 g Bacto peptone; 15 g Bacto agar; 30 mg streptomycin sulfate; 250 mg ampicillin; 10 mg rifampicin; 500mg pentachloronitrobenzene; 500 mg dicloran; and 1 L distilled water) was superior to potato dextrose agar in detecting C. destuctans in diseased roots. Isolation of C. destructans from diseased seedlings arising from seeds sown in replant soil supported the hypothesis that this pathogen is a cause of ginseng replant failure in North America.

Aspergillus nidulans 온도감수성 돌연변이주의 분리 및 분석 (Isolation and Analysis of Temperature Sensitive Mutants in Aspergillus nidulans)

  • 박찬규;강현삼
    • 미생물학회지
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    • 제16권3호
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    • pp.111-121
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    • 1978
  • About 40 temperature-sensitive mutants have been isolated as a preliminary step to study the spore germination, the cell cycle, and the control of macromolecular synthesis in Aspergillus nidulans. To obtain temperature-sensitive mutants rapidly and effectively, the selective enrichment method using antifungal antibiotic nystatin was developed. Based on the data which had applied to the concentration of auxotrophic mutants by the earlier investigators, the optimal concentration and the time of treatment at the nonpermissive temeprature were determined as 50 to 100 units per ml and 4.5 hr., respectively. Out of 41 ts mutants assigned to the strain symbol PK, thirteen that seemed to be arrested at the earlystage of spore germination were subjected to the further cytological and genetic analysis. Elght of these mutants are able to form germ tube and five not. Staining with acid fuchsin for the 5PK strains shows that one irreversible mutant, PK6 strain able to form germ tube, accumulate mitotic spindle, being arrested in mitosis. Another PK15 and PK23 strain have more than one intact nucleolus without germ tube formation at the restrictive temperature. the temperature-senstive mutation in PK12 strain, the onlystrain which is able occurred in certain gene specific for the germination of spore. All of the ts markers are recessive and complement each other in heterokaryon between two different ts markers at the restrictive temperature.

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STI-CMP 공정 적용을 위한 연마 정지점 고찰 (A Study of End Point Detection Measurement for STI-CMP Applications)

  • 이경태;김상용;김창일;서용진;장의구
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2000년도 하계학술대회 논문집
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    • pp.90-93
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    • 2000
  • In this study, the rise throughput and the stability in fabrication of device can be obtained by applying of CMP process to STI structure in 0.18um semiconductor device. To employ in STI CMP, the reverse moat process has been added thus the process became complex and the defects were seriously increased. Removal rates of each thin films in STI CMP was not equal hence the devices must to be effected, that is, the damage was occured in the device dimension in the case of excessive CMP process and the nitride film was remained on the device dimension in the case of insufficient CMP process than these defects affect the device characteristics. To resolve these problems, the development of slurry for CMP with high removal rate and high selectivity between each thin films was studied then it can be prevent the reasons of many defects by reasons of many defects by simplification of process that directly apply CMP process to STI structure without the reverse moat pattern process.

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Isolation of Novel Pseudomonas diminuta KAC-1 Strain Producing Glutaryl 7-Aminocephalosporanic Acid Acylase

  • Kim, Dae-Weon;Kang, Sang-Mo;Yoon, Ki-Hong
    • Journal of Microbiology
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    • 제37권4호
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    • pp.200-205
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    • 1999
  • 7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.

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Raw Animal Meats as Potential Sources of Clostridium difficile in Al-Jouf, Saudi Arabia

  • Taha, Ahmed E.
    • 한국축산식품학회지
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    • 제41권5호
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    • pp.883-893
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    • 2021
  • Clostridium difficile present in feces of food animals may contaminate their meats and act as a potential source of C. difficile infection (CDI) to humans. C. difficile resistance to antibiotics, its production of toxins and spores play major roles in the pathogenesis of CDI. This is the first study to evaluate C. difficile prevalence in retail raw animal meats, its antibiotics susceptibilities and toxigenic activities in Al-Jouf, Saudi Arabia. Totally, 240 meat samples were tested. C. difficile was identified by standard microbiological and biochemical methods. Vitek-2 compact system confirmed C. difficile isolates were 15/240 (6.3%). Toxins A/B were not detected by Xpect C. difficile toxin A/B tests. Although all isolates were susceptible to vancomycin and metronidazole, variable degrees of reduced susceptibilities to moxifloxacin, clindamycin or tetracycline antibiotics were detected by Epsilon tests. C. difficile strains with reduced susceptibility to antibiotics should be investigated. Variability between the worldwide reported C. difficile contamination levels could be due to absence of a gold standard procedure for its isolation. Establishment of a unified testing algorithm for C. difficile detection in food products is definitely essential to evaluate the inter-regional variation in its prevalence on national and international levels. Proper use of antimicrobials during animal husbandry is crucial to control the selective drug pressure on C. difficile strains associated with food animals. Investigating the protective or pathogenic potential of non-toxigenic C. difficile strains and the possibility of gene transfer from certain toxigenic/ antibiotics-resistant to non-toxigenic/antibiotics-sensitive strains, respectively, should be worthy of attention.