• Journal of Microbiology and Biotechnology
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• 제4권3호
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• pp.176-182
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• 1994

The light-organ symbiont of pine cone fish, Vibrio fischeri, senses its presence in the host and responds to environmental changes by differentially expressing its symbiosis-related luminescence genes. The V. fischeri luminescence genes are activated by LuxR protein in the presence of an autoinducer. In an effort to elucidate the mechanism of regulation of luxR, a plasmid containing luxR was mutagenized in vitro with hydroxylamine and a luxR mutant plasmid was isolated by its ability to activate luminescence genes cloned in E. coli in the absence of the autoinducer. The specific base change identified by DNA sequencing was only single base transition at ＋78 from the transcriptional start of luxR. Based on a Western immunoblot analysis, the nucleotide change directed the synthesis of much higher level of LuxR protein without any amino acid substitutions. The results suggest that the region including the ＋78th base is presumably internal operator required for autorepression of luxR, and the increased cellular level of LuxR results in activation of luminescence genes by autoinducer independent fashion.

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1197-1206
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• 2005

Vibrio vulnificus is an opportunistic pathogen that causes a septicemia and expresses numerous virulence factors, in which luxS and smcR are genes encoding for components responsible for quorum-sensing regulation. In the present study, null mutants were constructed with lesions in each or both of these two genes from the V. vulnificus Vv$\Delta$Z strain, which is a lacZ$^{-}$ and chloramphenicol/streptomycin-resistant derivative of the wild-type ATCC29307 strain, and their phenotypes related to virulence were compared with those of the parental cells. $LD_{50}$ and histopathological findings of luxS-, smcR-, or luxS- smcR- deficient mutant were not different from those of the parent strain, a lacZ-deficient streptomycin-resistant strain in mice. However, time of death in mice was delayed, and numbers of bacteria survived in bloodstream after intraperitoneal injection in mice were decreased by mutation, especially luxS and smcR double mutant (VvSR$\Delta$ZSR). These phenomena were supported by increased serum sensitivity and delayed bacterial proliferation in both murine blood and iron-restricted medium. These results suggest that the luxS and luxR homologous genes in V. vulnificus could playa role in bacterial survival in host by enhancing proliferation and adjusting to changed environment.

• 한국작물학회지
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• 제32권3호
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• pp.256-267
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• 1987

포장상태에서 인삼생육의 최적광양을 알기 위하여 상대조도 5%(관행볏짚). 15 % 및 20 % (백색불섬포) 하에서 5년근 개체군의 재식위치별로 해가 림내 조도, 미기상, 광합성 및 호흡속도, 근수량등을 조사한 바 그 결과는 다음과 같다. 1. 해가림 내 맑은날의 온도는 10~14시경에 상대조도 5 %구에 비해서 15%구가 2$^{\circ}C$, 20 %구가 3$^{\circ}C$정도 각각 높았으나 흐린날은 상대조도 처리간에 차이가 거의 없었다. 2. 해가림내 맑은날 상대습도는 10~14시경 상대조도 5%구에 비해서 15 %구가 5%, 20 %구가 8%정도 각각 낮았고, 흐린날은 차이가 근소하였다. 3. 해가림내 맑은날의 일중 조도는 10~15시경에 상대조도 5%구는 5,000 lux 미만이었으나 15%구는 15,000 lux, 20%구는 20,000 lux로 이논치와 비슷하였으나, 흐린날은 조도의 변화폭이 심하여 대개 상대조도 5%구는 3,0001ux 미만이었으나 15 % 및 20%고는 각각 10,000 lux, 15,000 lux 였다. 4. 포장하의 광합성속도는 맑은날의 경우 상대조도 15% 및 20%구가 5%구에 비해 현저히 빨랐고, 흐린날의 경우도 비슷하였다. 5. 일중 광합성 총양은 맑은날과 흐린날 공히 상대조도 15%구가 가장 높고 20%, 5% 순으로 높았으며. 맑은날이 흐린날보다 광합성량이 많았다. 행간 광합성량은 상대조도 15% 및 20%구는 비슷한 반면 5%구는 차이가 컸다. 6. 근중은 15%, 20%. 5% 순으로 상대조도 15%구가 가장 무거웠고 행간의 근중차이가 15%, 20%구는 비슷한 반면 5%구는 컸다.

• Journal of Microbiology and Biotechnology
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• 제19권5호
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• pp.431-438
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• 2009

The alkaline serine protease asp, which was shown to be a virulence factor of Vibrio alginolyticus as a purified protein, was cloned from V. alginolyticus EPGS, a strain recently isolated from moribund Epinephelus coioides in an outbreak of vibriosis in a mariculture farm of Shenzhen. The asp null mutant was constructed by homologous recombination with suicide plasmid pNQ705-1. Compared with the wild-type strain, the asp null mutant exhibited a significant decrease of total extracellular protease activity, and caused a IS-fold decrease in virulence of V. alginolyticus. In our previous study, the luxO and $luxR_{val}$ genes from V. alginolyticus MVP01 were cloned and identified, and the luxO-$luxR_{val}$ regulatory couple was shown to regulate various genes expression, suggesting that it played a central role in the quorum sensing system of V. alginolyticus. In this study, the regulation of the asp gene was analyzed by using RT-PCR and quantitative real-time PCR methods; we proved that its transcription was greatly induced at the late stage of growth and was regulated by a luxO-$luxR_{val}$ regulatory system.

• Journal of Microbiology
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• 제38권2호
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• pp.80-87
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• 2000

The nucleotide sequence of the luxC gene coding for lux-specific fatty acyl-CoA reductase and the upstream DNA (325bp)of the structural gene from bioluminescent bacterium, Photobacterium phosphoreum, has been deternubed. An open reading frame extending for more than 20 codons in 325 bp DNA upstream of luxC was not present in both directions. The lux gene can be translated into a polypeptide of 54 kDa and the amino acid sequences of lux specific reductases of P. phosphoreum shares 80, 65, 58, and 62% identity with those of the Photobacterium leiognathi, Vibrio fischeri, Vibrio harveyi, and Xehnorhabdus luminescenens reductases, respectively. Analyses of codon usage, showing that a high frequency (2.3%) of the isoleucine codon, AUA, in the luxC gene compared to that found in Escherichia coli genes (0.2%) and its absence in the luxA and B genes, suggested that the AUA codon may play a modulator role in the expression of lux gene in E. coli. The structural genes (luxC, D, A, B, E) of the P. phosphoreum coding for luciferase (${\alpha}$,${\beta}$) and fatty acid reductase (r, s, t) polypeptides can be expressed exclusively in E. coli under the T7 phage RNA polymerase/promoter system and identificationof the [35S]methionine labelled polypeptide products. The degree of expression of lux genes in analyses of codon usage. High expression of the luxC gene could only be accomplished in a mutant E. coli 43R. Even in crude extracts, the acylated acyl-CoA reductase intermediate as well as acyl-CoA reductrase activities could be readily detected.

• Journal of Microbiology and Biotechnology
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• 제19권8호
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• pp.765-773
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• 2009

Edwardsiella tarda is a pathogen with a broad host range that includes human and animals. The E. tarda hemolysin (Eth) system, which comprises EthA and EthB, is a noted virulence element that is widely distributed in pathogenic isolates of E. tarda. Previous study has shown that the expression of ethB is regulated by iron, which suggests the possibility that the ferric uptake regulator (Fur) is involved in the regulation of ethB. The work presented in this report supports the previous findings and demonstrates that ethB expression was decreased under conditions when the E. tarda Fur ($Fur_{Et}$) was overproduced, and enhanced when $Fur_{Et}$ was inactivated. We also identified a second ethB regulator, EthR, which is a transcription regulator of the GntR family. EthR represses ethB expression by direct interaction with the ethB promoter region. In addition to ethB, EthR also modulates, but positively, luxS expression and AI-2 production by binding to the luxS promoter region. The expression of ethR itself is subject to negative autoregulation; interference with this regulation by overexpressing ethR during the process of infection caused (i) drastic changes in ethB and luxS expressions, (ii) vitiation in the tissue dissemination and survival ability of the bacterium, and (iii) significant attenuation of the overall bacterial virulence. These results not only provide new insights into the regulation mechanisms of the Eth hemolysin and LuxS/AI-2 quorum sensing systems but also highlight the importance of these systems in bacterial virulence.

• 미생물학회지
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• 제45권4호
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• pp.419-424
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• 2009

발광 세균 Photobacterium과 Vibrio의 lux 유전자가 삽입된 재조합 플라스미드를 유전자 전이시킨 발광표현형 대장균이 내는 빛의 세기를 조사하였다. 여러 대장균 균주에 형질전환 시켰는 바, 빛을 내는데 관여하는 효소들을 코드하는 유전자를 모두 포함하는 Photobacterium leiognathi lux 오페론이 삽입된 재조합플라스미드(PlXba.pT7-3)가 형질전환된 대장균 43R (Escherichia coli 43R) 균주에서는 발광세기가 다른 균주에 비해 1,000배 이상 되었으며, Vibrio harveyi luxA 유전자와 luxB 유전자를 융합시킨 유전자가 삽입된 재조합플라스미드(VhluxAB.pT7-5)가 형질전환 된 대장균 43R (E. coli 43R)에서는 기질인 fatty aldehyde를 가하면 단일 콜로니에서도 빛을 볼 수 있게 되는 대장균 형질전환체를 얻었다. 또한 이들이 중금속에 노출되었을 때 생물발광이 감소하였으며, 이들 발광 대장균 형질 전환체가 담긴 고정화된 세포에서도 빛을 내는 것을 관측함으로써 이들 실험 결과들이 lux 유전자를 활용한 바이오센서 시스템 개발의 기반 기술이 될 가능성을 보였다.

• Interdisciplinary Bio Central
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• 제3권3호
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• pp.10.1-10.8
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• 2011

Introduction: Hash functions are computer algorithms that protect information and secure transactions. In response to the NIST's "International Call for Hash Function", we developed a biological hash function using the computing capabilities of bacteria. We designed a DNA-based XOR logic gate that allows bacterial colonies arranged in a series on an agar plate to perform hash function calculations. Results and Discussion: In order to provide each colony with adequate time to process inputs and perform XOR logic, we designed and successfully demonstrated a system for time-delayed bacterial growth. Our system is based on the diffusion of ${\ss}$-lactamase, resulting in destruction of ampicillin. Our DNA-based XOR logic gate design is based on the op-position of two promoters. Our results showed that $P_{lux}$ and $P_{OmpC}$ functioned as expected individually, but $P_{lux}$ did not behave as expected in the XOR construct. Our data showed that, contrary to literature reports, the $P_{lux}$ promoter is bidirectional. In the absence of the 3OC6 inducer, the LuxR activator can bind to the $P_{lux}$ promoter and induce backwards transcription. Conclusion and Prospects: Our system of time delayed bacterial growth allows for the successive processing of a bacterial hash function, and is expected to have utility in other synthetic biology applications. While testing our DNA-based XOR logic gate, we uncovered a novel function of $P_{lux}$. In the absence of autoinducer 3OC6, LuxR binds to $P_{lux}$ and activates backwards transcription. This result advances basic research and has important implications for the widespread use of the $P_{lux}$ promoter.

• 감성과학
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• 제18권2호
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• pp.45-54
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• 2015

본 연구에서는 LED 광원에 대하여 재실자의 업무 효율성과 에너지 저감 효과를 알아보기 위한 실험을 실시하였다. 이를 위해 LED 광원의 특성인 펄 스 폭변조(PWM, pulse width modulation)와 조도(lux)를 제어하여 총 9가지의 다양한 조명환경을 구성하였다. LED 조명의 펄스변조 비율은 각각 R:G:B=1:1:1, R:G:B=4:1:5, R:G:B=8:7:7으로 하였으며, 조도는 각각 400 lx, 700 lx, 1000 lx 으로 설정하였다. 또한, 실내환경은 온도 $20{\sim}24^{\circ}C$, 습도 50~60%, 착의량 1 clo 로 설정하였다. 각각의 주어진 9개의 조명환경에서 업무 효율성과 에너지 소비에 대해 분석하였다. 업무 효율성 분석을 위해 오류검색수정 작업을 실시하였으며, 에너지 소비 분석을 위해 각 조명환경에서 누적 소비전력을 측정하였다. 제안한 조명환경을 통해 실험한 결과, 업무 효율성은 400 lx 보다 700 lx 이상에서 정확도 및 소요시간의 효율이 좋았으며, 소요시간의 경우 제안한 펄스변조 R:G:B=8:7:7 에서 가장 좋은 효율을 나타냈다. 또한, 각각의 조도에 대한 소비전력은 펄스변조 R:G:B=8:7:7 > RGB=1:1:1 > R:G:B=4:1:5의 순으로 낮게 나타났다. 따라서, 본 논문에서 제안한 펄스 폭 변조 효과가 업무효율성 및 에너지 저감에 영향을 미치는 것을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.219-227
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• 2008

The free-living photoheterotrophic Gram-negative bacterium Rhodobacter sphaeroides possesses a quorum-sensing (QS) regulatory system mediated by CerR-CerI, a member of the LuxR-LuxI family. To identify the genes affected by the regulatory system, random lacZ fusions were generated in the genome of R. sphaeroides strain 2.4.1 using a promoter-trapping vector, pSG2. About 20,000 clones were screened and 23 showed a significantly different level of ${\beta}$-gal activities upon the addition of synthetic 7,8-cis-N-tetradecenoyl-homoserine lactone (RAI). Among these 23 clones, the clone showing the highest level of induction was selected for further study, where about a ten-fold increase of ${\beta}$-gal activity was exhibited in the presence of RAI and induction was shown to be required for cerR. In this clone, the lacZ reporter was inserted in a putative gene that exhibited a low homology with catD. A genetic analysis showed that the expression of the catD homolog was initiated from a promoter of another gene present upstream of the catD. This upstream gene showed a strong homology with luxR and hence was named qsrR (quorum-sensing regulation regulator). A comparison of the total protein expression profiles for the wild-type cells and qsrR-null mutant cells using two-dimensional gel electrophoresis and a MALDI-TOF analysis allowed the identification of sets of genes modulated by the luxR homolog.