• Title/Summary/Keyword: 형광성 박테리아

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Motion of Microbeads Propelled by Bacterial Chemotaxis (박테리아의 주화성에 의한 미세입자의 운동)

  • Kim, Dong-Wook;Kim, Young-Won;Yoo, Jung-Yul
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.34 no.5
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    • pp.523-529
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    • 2010
  • Recently, several research groups have been investigating the motion of flagellated bacteria, with the aim of examining the feasibility of using bacterial chemotaxis as an efficient power source for microactuators. In this study, microparticle-tracking velocimetry ($\mu$-PTV) is used for investigating the motion of fluorescent microbeads propelled by bacterial chemotaxis. Flagellated bacteria, Serratia marcescens, are spontaneously attached to the surface of the fluorescent polystyrene (PS) microbeads in an aqueous culture. The microbeads thus treated are injected into the test medium, which contains the solidified chemoattractant L-aspartate. With time, the particles slowly move toward the zone in which the L-aspartate concentration is high. This study shows that chemotaxis of flagellated bacteria can be applied as an efficient power source for microactuators.

Site-specific Dye-labeling of the Bacterial Cell Surface by Bioconjugation and Self-assembly (바이오접합과 자가결합을 이용한 박테리아 세포막의 위치 특이적 형광 표지)

  • Yang, I Ji;Lim, Sung In
    • Korean Chemical Engineering Research
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    • v.60 no.3
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    • pp.398-406
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    • 2022
  • The outer membrane of Gram-negative bacteria is the outermost layer of cellular environment in which numerous biophysical and biochemical processes are in action sustaining viability. Advances in cell engineering enable modification of bacterial genetic information that subsequently alters membrane physiology to adapt bacteria to specific purposes. Surface display of a functional molecule on the outer membranes is one of strategies that directs host cells to respond to a specific extracellular matter or stimulus. While intracellular expression of a functional peptide or protein fused to a membrane-anchoring motif is commonly practiced for surface display, the method is not readily applicable to exogenous or large proteins inexpressible in bacteria. Chemical conjugation at reactive groups naturally occurring on the membrane might be an alternative, but often compromises fitness due to non-specific modification of essential components. Herein, we demonstrated two distinct approaches that enable site-specific decoration of the outer membrane with a fluorescent agent in Escherichia coli. An unnatural amino acid genetically incorporated in a surface-exposed peptide could act as a chemoselective handle for bioorthogonal dye labeling. A surface-displayed α-helical domain originating from a part of a selected heterodimeric coiled-coil complex could recruit and anchor a green fluorescent protein tagged with a complementary α-helical domain to the membrane surface in a site- and hetero-specific manner. These methods hold a promise as on-demand tools to confer new functionalities on the bacterial membranes.

Toxicity Evaluation of Tar Colors by Water Fleas and Luminescent Bacteria (물벼룩과 형광성 박테리아를 이용한 타르색소의 독성평가)

  • Choo, Yeon Jong;Kim, Gun Heung;Kim, Hung Soo
    • Journal of Wetlands Research
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    • v.9 no.1
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    • pp.21-29
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    • 2007
  • In Korea, we assign the chemical substances of 535 types as toxic substance. Only 10% of the 535 toxic substances are being managed by the Ministry of Environment related with water quality standard. Tar color is also one of chemical substances, but we have the lacks for the information of tar colors about the environmental effects of aquatic ecosystem. This study performed the test of bioassay using Water Fleas and Luminescent Bacteria. The tar has 7 types of colors allowed as the edible color and we evaluate the toxicities of 5 tar colors out of 7 colors and we would like to provide the informations for further study as we perform the toxicity test for the samples of 5 tar colors. We did the toxicity test of using Water Fleas From the results, we obtained the magnitudes of toxicity in order of Red No.2, Yellow No.5, Red No.3, Yellow No.4, Blue No.1. As the result based on Microtox Acute Toxicity Test using Luminescent Bacteria with the standard of 15min-EC50, we obtained in order of Yellow No.5, Food Red No.3, Red No.2, Yellow No.4, Blue No.1. We could expect the tar colors may have different effects on the aquatic ecosystem, respectively and it may influence to the aquatic ecosystem and the human, because of bioconcentration by food chain when toxicity of the tar colors overflow in the aquatic ecosystem.

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Microcontact Printing of Bacteria Using Hybrid Agarose Gel Stamp (혼성 아가로즈젤 스탬프를 이용한 박테리아 마이크로 컨택트 프린팅)

  • Shim, Hyun-Woo;Lee, Ji-Hye;Lee, Chang-Soo
    • KSBB Journal
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    • v.21 no.4
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    • pp.273-278
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    • 2006
  • The noble method of hybrid agarose gel microstamp fabricated by replica molding against PDMS master to make bacteria patterns on agar surface was presented. After the fabricated hybrid agarose gel microstamp was inked with E. coli, we could obtain 2 dimensional bacterial arrays with $50{\mu}m$ circular spots. And the various shaped patterns based on experimental design were easily generated. The analysis of mean fluorescent signal was showed that bacterial pattern have high contrast between spots and background and homogeneity of pattern. Our proposed method solved the problem of wetting and handling with small soft agarose gel microstamp when bacteria were used for ink. The agarose gel stamp provides appropriate environment to inked bacteria, which is essential technology for cell patterning with high retaining viability during the patterning process. This method is reproducible, convenient, rapid, and could be applied to screening system, study of cell-surface interaction, and microbial ecology.

Acute Toxicity on Daphnia magna and Photobacterium phosphoreum for synthetic Detergents (물벼룩과 형광성 박테리아를 이용한 합성세제의 급성독성평가)

  • 김태영;채수권;김건흥
    • Water for future
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    • v.27 no.1
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    • pp.69-77
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    • 1994
  • As the standard of living improves, the amount of synthetic detergent consumption greatly increases. Detergents which are not treated in the sewer treatment processes, flow into rivers or waterstreams and accelerate the pollution of the surface water resources. Detergents contain lots of toxicants. And it is difficult to evaluate gross toxicity of each toxicant in thereceiving water. In the study, the acute toxicity of the synthetic detergents for home laundering and kitchen use were monitored with daphnia magna and photobacterium phosphoreum. Seven kinds of detergents were tested to evaluate the acute toxicity. The mean 24hr, 48hr-LC50 of the synthetic detergent for home laundering were 4.25%, 2.50% and these for kitchen use were 2.01%, 1.36% respectively. And the mean 5min, 15min-EC50 of the synthetic detergent for home laundering were 1.83%, 1.02%.

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An Experimental Study on Toxicity Evaluation of Melting Slag from MSWI Ash Using Microtox Bioassay (Microtox 생물검정을 이용한 소각재 용융슬래그의 독성평가)

  • Park, Sang-Goo;Kim, Geon-Hung;Han, Yang-Soo;Kim, Gil-Ho
    • Proceedings of the Korea Water Resources Association Conference
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    • 2012.05a
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    • pp.674-677
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    • 2012
  • 본 연구에서는 기존 매립이나 지반보강재로 사용되던 소각재 용융슬래그를 고부가가치 수처리 여재로 활용하기 위하여 형광성 박테리아를 통한 Microtox 생물검정법으로 독성을 평가하였다. 소각재 용융슬래그의 독성평가 대조군으로는 기존의 수처리 여재인 입상활성탄(석탄계/야자계/목탄계), PP PE펠렛(Poly-Propylene Poly-Ethylene pallet), 표준여과사를 비교하였으며, 시료의 용출시험은 US EPA에서 제안한 TCLP 방법을 사용하였다. Microtox Acute Toxicity 평가 결과, 독성순위는 석탄계 입상 활성탄, 소각재 용융슬래그, 목탄계 입상활성탄, 야자계 입상활성탄, 표준여과사, PP PE 순으로 나타났으며, 실험에 사용된 모든 수처리 여재들은 급성독성을 고려하지 않는 무독성으로 나타났다. 따라서 소각재 용융슬래그가 수처리를 목적으로 수체에 편입시켜도 기존의 수처리 여재들과 비교할 때 수환경에 미치는 영향은 미미할 것으로 판단된다.

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광학 분석 시스템용 수.발광 소자 집적 모듈 개발

  • Song, Hong-Ju;Lee, Jun-Ho;Park, Jong-Hwan;Han, Cheol-Gu;Park, Jeong-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.08a
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    • pp.311-312
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    • 2012
  • 본 발표에서는 광학적 분석 시스템에 적용 가능한 발광소자(광원)과 수광소자(광센서)를 집적화시키는 모듈(수 발광 집적모듈) 기술을 제시하고자 한다. 이러한 수-발광 집적모듈은 다양한 응용 분야에 적용 될 수 있다. 예를 들어, 광신호 감지를 위한 광통신용 송-수신 모듈(optical communication), 의료/진단 분야에서 단백질/DNA/박테리아 등의 검출 및 분석에 관한 바이오 센서(bio-sensor), 그리고 대기(가스)/수질 모니터링에 관한 환경센서 등 매우 광범위한 분야에 해당되는 요소 기술이라 할 수 있다. 특히, 이들 분야들 중 바이오 물질을 분석하고 검출하는 광학적 바이오 센서 기술은 높은 경제적 가치와 산업적 성장 잠재력으로 인해 오랫동안 활발한 연구가 진행되어 오고 있다. 이러한 광학적 바이오 센서에서 가장 범용적인 방법 중 하나가 항온-항체 면역반응을 기반으로 하는 형광 검출(fluorescence detection) 기법이다. 이러한 시스템은 전체적으로 광원, 광학계, 그리고 센서로 구성되는데 기존에 일반적으로 사용되고 있는 형광 현미경의 경우는 민감도가 우수하다는 장점은 있으나 상당히 고가이고 부피가 크며 복잡한 광학구성으로 이루어져 있다는 한계점을 가지고 있다. 이러한 맥락에서 고민감도를 확보하면서 휴대성, 고속처리, 저가 등의 특성을 가진 시스템에 대한 요구가 갈수록 증가하고 있다. 이를 해결하기 위한 핵심기술 중의 하나가 수-발광 부분을 집적화 시키는 기술이라 할 수 있다. 본 연구에서는 바이오 센서 기술의 하나로서 형광을 측정하여 혈액내의 진단 지표인자를 검출할 수 있는 휴대용 혈액진단기기에 적용되는 소형 수 발광 집적 모듈을 개발하였다. 혈액내의 검출 성분의 양에 따라 형광의 세기가 변화하게 됨으로써 정량적인 검출이 가능한 원리이다. 모듈의 구조는 크게 광원(발광소자), 광학계, 그리고 광센서(수광소자) 세 영역으로 나누어 진다. 광원은 635 nm 적색 레이저다이오드로서 형광체(Alexa Fluor 647/발광파장: 668 nm)를 여기 시키는 기능을 하며 장착된 볼렌즈 의해 샘플의 형광체 영역으로 집광된다. 광학계는 크게 시준렌즈(collimating lens)와 광학필터로 구성됨으로써 샘플로부터 발생되는 광을 적절하게 수광소자로 전달하는 기능을 하게 된다. 여기서 광학필터의 경우는 기본적으로 Distributed Bragg's Reflector(DBR) 구조로써 실리콘(Si) 포토다이오드 상부에 모노리식(monolithic)하게 형성되며 검출 샘플로부터 진행되는 레이저 광(잡음의 주원인)은 차단하고 형광(광신호)만 통과 시키는 기능을 하게 된다. 따라서 신호 대 잡음비(S/N ratio)를 향상시키기 위해서는 정밀한 광 필터링 기능이 요구됨으로써 박막의 세밀한 공정 조건과 구조적-광학적 특성 분석이 수행되었다. 마지막으로 포토다이오드 소자는 일반적인 구조 이외에 중앙에 원형 구멍이 형성된 특별한 구조가 적용된다. 이것은 포토다이오드 구조에 변화를 줌으로써 모듈 구조를 효율적으로 응용할 수 있다는 의미를 갖는다. 또한 포토다이오드의 전기적-광학적 측정 분석을 통해 잡음 및 감도 특성이 세부적으로 조사되며 형광신호를 효과적으로 측정할 수 있음을 확인하였다. 최종적으로 제작된 모듈은 약 $1{\times}1{\times}1cm^3$ 내외 정도의 크기를 갖는다. 요약하자면 본 발표에서는 광학적 바이오센서에 적용할 수 있는 소형 수-발광 소자 집적모듈을 소개한다. 전체 모듈 설계는 최소한의 부피를 가짐과 동시에 측정의 정밀성을 향상시키는데 초점을 맞추어 진행하였다. 세부요소인 광학필터와 포트다이오드의 경우 잡음 및 민감도에 미치는 중요성 때문에 세밀한 공정 및 특성분석이 수행되었다. 결론적으로 독자적인 설계 및 공정을 통해 휴대성 및 정밀성 등의 목적에 부합한 경쟁력 있는 수-발광 소자 집적모듈 제작 기술을 확보하였다.

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Utilization of Waste Tires as Soil Reinforcement; (2) Environmental Effects (지반보강재로서 폐타이어의 활용; (2) 환경적 영향)

  • 윤여원;문창만;김건흥
    • Journal of the Korean Geotechnical Society
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    • v.20 no.3
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    • pp.119-128
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    • 2004
  • Environmental impact of waste tires as gound-reinforcing material is studied. Analysis for chemical compounds and toxic effect were performed on effluents from twelve lysimeters in which waste tires were mixed with sand and three initially different environmental solutions of acidic, neutral, and basic circulated through the mixture. The test results of effluents collected from the lysimeters provided that the contaminant concentrations were lower than those of Korean drinking water standards for all the selected and tested metal elements. While iron concentration increased slightly with the exposure period, other metal concentrations decreased with the number of circulation times. From the comparison with previous investigations, the contaminant concentration decreased with the increase of tire size, i.e. increases with the increase of the exposed surface of tire metals. From the toxicity tests, no deteriorative effect was observed and it could be concluded that waste tires are not biologically hostile materials.

Generation and Expression of Amino-Terminal Domain of the Gene Coding for the Lumazine Protein from Photobacterium phosphoreum (발광 박테리아 Photobacterium phosphoreum의 Lumazine Protein을 코드 하는 유전자의 염기 서열 분석 및 발현)

  • Woo Young-Eun;Kim So-Young;Lee Chan-Yong
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.306-311
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    • 2005
  • In this study, the amino-terminal half truncated lump and the whole lump genes from Photobacterium phosphoreum coding for the lumazine protein were cloned by polymerase chain reaction and expressed in Escherichia coli. To identifiy of the binding site of the ligand or substrate, the amino acid identities from the sequences of the lumazine protein, yellow fluorescent protein, and riboflavin synthase from different organisms were also compared and analyzed.

The Electrochemical Chlorination for Marine Plankton Community Disinfection (해양 플랑크톤 군집의 전기분해 염소소독 효과)

  • Kang, Jung-Hoon;Shin, Kyoung-Soon;Hyun, Bong-Gil;Jang, Min-Chul;Kim, Eun-Chan;Chang, Man
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.10 no.3
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    • pp.127-137
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    • 2007
  • To confirm whether or not the Electrochemical Disinfection System (EDS) meet with the D-2 regulation established by IMO (International Maritime Organization), the biological treatment efficacy of the EDS was assessed using three groups of natural marine plankton (bacteria, $10-50\;{\mu}m$ and $>50\;{\mu}m$ sized organisms). Influent water was passed through the EDS under the flow velocity ($23.8\;m^3/hr$) and test design was consisted of control (no treatment) and experimental (10 ppm and 30 ppm) condition for total residual chlorine (TRC). And the biological condition of the influent water followed the standards established by the guidelines for the approval of ballast water management systems. The disinfection efficacy of the $10-50\;{\mu}m$ sized organisms (phytoplankton) was assessed by three kinds of measurements using photomicroscope, epifluorescence microscope and fluorometer (fumer Designs 10-AU). After being passed through the EDS, all motile phytoplankton lost their motility under photomicroscope, the colour of chlorophyll fluorescence fumed from red into green under epifluorescence, and the high chlorophyll fluorescence (Expt. 1: 6.95, Expt. 2: 7.11) detected by fluorometer decreased into value not detected. These results indicated phytoplankton community was totally killed after electrochemical disinfection treatment. Survivorship of the larger organisms than $50\;{\mu}m$ was determined based on the appendage's movement under a stereomicroscope. Natural assemblage collected from ambient seawater was killed shortly after being passed through the EDS, whereas some Artemia remained alive. However, no live Artemia was found after 24 hour further exposure to each TRC concentration (10 and 30 ppm) under darkness. After electrochemical treatment, the target bacteria such as aerobes, coliform and Escherichia coli were completely killed on the basis of CFU (colony forming unit) on Petrifilm plate ($3\;M^{TM}$) after 48 hr incubation. Moreover, no regrowth was found in the three groups of plankton during five days under additional exposure to the treated water. These results indicated that the disinfection efficiency of the EDS on the three groups of plankton satisfy D-2 regulation.

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