• 대한환경공학회지
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• 제27권5호
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• pp.507-512
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• 2005

본 연구에서는 유전자 재조합 발광균주, Pseudomonas putida mt-2 KG1206을 이용하여 토양에 오염된 m-toluate 탐지 방법 및 적용 가능성에 대해 조사하였다. KG1206은 톨루엔 계열 화합물의 중요 중간 분해물질인 m-toluate 및 benzoate가 직접 생물 발광 유도제로 작용하며, 또한 톨루엔 계열 화합물들이 간접 유도제로서 발광 활성을 나타내었다. 토양에 오염된 유도제 오염원 검출을 위해 발광 균주 9.9 mL에 에탄올 추출물 0.1 mL을 첨가하여 조사하였다. 생물발광에 근거하여 작성된 m-toluate 검량선은 대략 $R^2>0.97$ 이상의 상관관계가 관찰되었다. 토양에 임의 오염된 m-toluate(직접 발광유도제)는 정립한 방법에 따라 발광활성에 근거하여 추측하였고, 기기분석치와 통계적으로 유의한 것으로 조사되었다. 본 연구 결과를 통해서 특정 화합물에 대해 발광을 생산하는 유전자 재조합 균주가 특정 오염원에 오염된 지역의 관리를 위한 수단으로 사용할 수 있는 가능성을 확인할 수 있었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.479-479
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• 2000

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.480-480
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• 2000

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 1991년도 춘계학술발표대회 논문집
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• pp.209-212
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• 1991

• 분석과학
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• 제23권1호
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• pp.60-67
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• 2010

본 연구는 생물발광단백질인 aequorin을 표지물질로 사용해 신경전달물질인 serotonin에 대한 생물 발광면역분석법을 처음으로 개발하였다: serotonin-avidin 접합체는 Mannich 법으로 합성하였으며, 최적화된 avidin: formaldehyde: serotonin의 초기 몰 비는 1:12,000:25 이었다. 개발된 serotonin에 대한 생물발광면 역분석법은 좋은 감도(LOD of 0.68 ng/mL)와 넓은 검출 범위($5.0{\times}10^{-10}\;M\sim5.0{\times}10^{-7}\;M$)를 나타내었다(참고, 혈청 내 serotonin의 양은 $151{\pm}45\;ng$/mL 이다.). 교차반응성에 대한 연구 결과, 5-methoxytryptamine은 28.0% 의 교차반응성을 보인 반면, 3-methylindole, melatonin 그리고 5-hydroxylindole-3-acetic acid 은 교차반응성을 보이지 않았다. 또한, 혈청에서의 회수율은 만족할만한 값이 얻어졌다. 그러므로, 본 연구에서 제안된 생물발광면역분석법은 혈청 내의 serotonin 을 모니터링 하는데 좋은 분석법으로 사용될 수 있다.

• 자원환경지질
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• 제49권5호
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• pp.411-415
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• 2016

환경독성법 (세균발광, 씨앗발아)을 이용하여 국내 벤토나이트 독성에 관한 기초 연구를 수행하였다. 생물발광과 상추를 이용한 씨앗발아에서의 독성범위는 각 -3 ~ 95%와 7.1 ~ 92.1%로 나타났다. 씨앗발아의 경우 상추 이외에 조사한 알타리무, 아욱의 씨앗 실험에서도 비슷한 결과를 나타내었다. 다양한 약산농도에서 추출한 점토광물이 함유하고 있는 중금속 함량과 독성의 상관관계는 거의 나타나지 않았다($R^2$ < 0.1). 특이한 사항은 점토광물의 색깔별 그룹에 따라 미치는 영향에 차이가 있음을 관찰할 수 있었다(청색 $67{\pm}8.7%$, 분홍 $27{\pm}10.4%$, 갈색 $58{\pm}14.5%$).

• Asian-Australasian Journal of Animal Sciences
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• 제11권5호
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• pp.498-502
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• 1998

The present study was designed, fIrst to develop the new methodology to measure the bioluminescence activity easily in live developing bovine embryos by photoncounting, and secondly to compare the expression efficiency of four luciferase reporter genes in bovine embryos at four- to 16-cell stages. In experiment 1, equimolar pSVlacZ and pSVEluc were microinjected into the pronucleus of fertilized bovine oocytes. At 2 days after micro injection, bioluminescence activity of these embryos was measured by photoncounting with a luminometer for 1 min, and lacZ gene expression in the same embryos was assayed by X-gal staining. All the luciferase-positive oocytes showed some bacterial ${\beta}$-galactosidase activity irrespective of the intensity. In experiment 2, four firefly luciferase genes (pTKEluc, pTK6WEluc, pSVEluc and pMiwluc) were introduced by micro injection, and the injected embryos were cultured for the following 2 days. Detection of the luciferase gene expression was done by photoncounting at 5 to 55 min. Over the measurement period, the luciferase activity was almost constant irrespective of the transgenes microinjected. The luciferase activity and expression efficiency at 2 days after microinjection were not significantly affected by the difference in the microinjected transgenes. The present results demonstrated that the bioluminescence activity in live developing bovine embryos could be measured quickly by photoncounting.

• Bulletin of the Korean Chemical Society
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• 제27권3호
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• pp.407-412
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• 2006

Bioluminescence single-site immunometric assay for methamphetamine (MA) using the native aequorin, a photoprotein, as a signal generator was developed for the first time. MA is a potent sympathomimetic amine with stimulant effects on the central nervous system. MA abuse induces hallucinations and, thus, may cause a serious social problem. The single-site immunometric MA assay was optimized and its dose-response behavior was examined. The dose-response curve shows that the detection limit is 1.1 ${\times}$ $10^{-10}$ M and a dynamic range is four orders of magnitude with 15 $\mu$g/mL BSA-MA conjugate and 1.0 ${\times}$ $10^{-8}$ M anti-MA antibody-biotin conjugate. In order to evaluate this assay, the structurally similar compounds, amphetamine, ephedrine, norephedrine, benzphetamine and N-4-(aminobutyl)methamphetamine were examined for their crossreactivity. None of these five compounds showed any cross-reactivity. Additionally, an artificial urine solution spiked with MA was analyzed by the MA assay, and the result of the analysis demonstrated the usefulness of the present assay for the determination of MA in urine.

• Journal of Microbiology
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• 제42권3호
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• pp.194-199
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• 2004

Investigation of the expression of the riboflavin (rib) genes, which are found immediately downstream of luxG in the lux operon in Photobacterium phosphoreum, provides more information relevant to the evolution of bioluminescence, as well as to the regulation of supply of flavin substrate for bacterial bioluminescence reactions. In order to answer the question of whether or not the transcriptions of lux and rib genes are integrated, a transcriptional termination assay was performed with P. phoxphoreum DNA, containing the possible stem-loop structures, located in the intergenic region of luxF and luxE ($\Omega$$\_$A/), of luxG and ribE ($\Omega$$\_$B/), and downstream of ribA ($\Omega$$\_$c/). The expression of the CAT (Chloram-phenicol Acetyl Transferase) reporter gene was remarkably decreased upon the insertion of the stem-loop structure ($\Omega$$\_$c/) into the strong lux promoter and the reporter gene. However, the insertion of the structure ($\Omega$$\_$B/) into the intergenic region of the lux and the rib genes caused no significant change in expression from the CAT gene. In addition, the single stranded DNA in the same region was protected by the P. phosphoreum mRNA from the Sl nuclease protection assay. These results suggest that lux genes and rib genes are part of the same operon in P. phosphoreum.

• Journal of Microbiology and Biotechnology
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• 제7권4호
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• pp.250-257
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• 1997

A new sensing system based on the immobilization of luminescent bacteria, photobacterium phosphoreum, was proposed for continuous real-time monitoring of pollutants. The response curves demonstrate that Photobacterium phosphoreum immobilized on the strontium alginate were very sensitive to seven reference chemicals used. The significant inhibitory concentrations for bioluminescence emission were 5 ppm for Pb$(NO_3)_2$), $NiCl_2$, $CdCl_2$, 50 ppm for $NaASO_2$, 0.1 ppm for $HgCl_2$, 0.5 ppm for pentachlorophenol and less than 5 ppm for SDS, respectively. The alginate mixed-cells (AMC) retained their luminescence during experimental period (29 days) under storage condition of $-80^{\circ}C$. The variables affecting performance of continuous flow through monitoring (CFTM) was optimized in order to ensure stability and efficiency. The flow through cell with strontium-alginate immobilized luminescent bacteria was tested with salicylate and 4-nitrophenol. A rapid response of luminescence was recorded by time drive mode in bioluminescence spectrometer after exposure to both toxicants.