• 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.

• 한국유기농업학회지
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• 제27권2호
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• pp.161-171
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• 2019

A light-emitting diode (LED) is most efficient in terms of economic benefits. The aim of the present study was to evaluate the effects of LED lighting intensity on milk production, milk composition, and the immune response of Holstein cows. Thirty-eight lactating cows were assigned to four experimental groups: control group (exposed natural daylight without LED light) and three LED treatment group (50 lux; natural daylight and 6 h LED light of 50 lux intensity, 100 lux; natural daylight and 6 h LED light of 100 lux intensity, 200 lux; natural daylight and 6 h LED light of 200 lux intensity). There was a significant effect on the decrease ratio in milk production in 200 lux group. Lactose was significantly increased in 100 lux than the other LED treated groups. In addition to the hemolytic biochemical analysis, AST and β-HB were significantly increased in the 50 lux than the other groups. Prolactin and IGF-1 levels were significantly increased in 200 lux compared to other groups. Besides, cortisol was significantly higher in 50 lux and lowered in the 200 lux than the control, while IgA and IgG were not significant among the groups. Therefore, it was concluded that LED lighting intensity had some impact on blood parameters and immune responses in dairy cows with increased milk production.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.101-105
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• 2003

Recombinant bioluminescent bacteria were used to monitor and classify the to xicity of azo dyes. Two constitutive bioluminescent bacteria, Photobacterium phosphoreum and Es-Cherichia coli, E, coli GC2 (lac::luxCOABE), were used to detect the cellular toxicity of the azo dyes. In addition, four stress-inducible bioluminestent E. coli, DPD2794 (recA::luxCDABE), a DNA damage Sensitive strain; DPD2540 (fabA::luxCDABE), a membrane damage sensitive strain; DPD2511 (katG::luxCDABE), an oxidative damage sensitive strain; and TV1061 (grpE::luxCDABE), a protein damage sensitive strain, were used to provide information about the type of toxicity caused by crystal violet, the most toxic dye of the 16 azo dyes tested. These results suggest that azo dyes result in serious cellular toxicity in bacteria, and that toxicity monitoring and classific ation of some azo dyes, In the field, may be possible using these recombinant bioluminescent bacteria.

• 미생물학회지
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• 제42권4호
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• pp.277-285
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• 2006

본 연구에서는 신경인성 방광으로 요도 카테터를 유치하고 있는 환자의 카테터로부터 카테터 내 요로감염(Catheter-Associate Urinary Tract Infection; CA-UTI)에 관여하는 박테리아인 Escherichia coli, Pseudomonas aeruginosa 그리고 Staphylococcus aureus를 순수 분리, 동정하였다. 이 균주들을 대상으로 하여 quorum sensing mechanism을 규명하는 기초 연구로 각 균주의 quorum sensing 신호물질인 autoinducer (AIs)를 합성하는 유전자의 mRNA 발현을 확인하고, 정략분석 하였다. 각 세 균주를 단일과 세 균주의 혼합으로 24시간, 30일 동안 배양하며 일정 시간 간격으로 sample을 얻었다. 이 중 24시간 배양한 sample을 가지고 reverse transcription polymerase chain reaction (RT-PCR)을 수행하여 각 AIs 합성 유전자가 발현되는 최초 박테리아 밀도를 확인하였다. 단일배양에서 E. coli와 S. aureus의 AIs 합성 유전자(ygaG와 luxS)의 mRNA가 발현되는 최초 박테리아 밀도는 $2.4{\times}20^5$ CFU/ml, $5.4{\times}10^6$ CFU/ml 이었으며 P. aeruginosa의 rhlI와 lasI의 경우 $6.9{\times}10^4$ CFU/ml로 나타났다. 세 균주의 혼합배양에서 ygaG와 luxS의 mRNA가 발현되는 최초 박테리아 밀도는$7.3{\times}10^5$ CFU/ml, $1.6{\times}10^7$ CFU/ml이었으며 rhlI와 lasI의 경우 $2.1{\times}10^5$ CFU/ml로 나타났다. 또한 30일 배양한 sample의 RT-PCR 결과, 배양초기부터 각 AIs 합성 유전자들의 mRNA가 30일 동안 일정한 양만큼 지속적으로 발현됨을 확인하였다. Real-time RT-PCR을 이용한 AIs 합성 유전자의 mRNA 발현을 정량 분석한 결과 각 균주에서 단일배양보다 혼합배양시 AIs 합성 유전자의 발현이 더 많았다. 가장 많은 발현량의 차이를 보인 경우 E. coli ygaG의 mRNA 발현량은 단일배양보다 세 균주의 혼합배양시 최고 약 30배 이상이 증가하였고, P. aeruginosa rhlI의 경우 단일배양보다 혼합배양시 최고 약 40배, P. aeruginosa lasI의 경우 최고 약 250배 그리고 S. aureus luxS의 경우는 단일배양보다 혼합배양시 최고 약 5배 이상 mRNA 발현량이 증가하였다. 또한 세 균주의 4가지 유전자 중 P. aeruginosa의 rhlI와 lasI의 mRNA가 가장 많은 양으로 발현됨을 확인하였다.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제29권7호
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• pp.1052-1059
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• 2016

Red light (RL) marked higher weight gain (WG) and preference of broilers compared to other light colors. This study aimed to investigate how different intensities of RL affect the performance, behavior and welfare of broilers. RL treatments were T1 = high intensity (320 lux), T2 = medium intensity (20 lux); T3 = dim intensity (5 lux), T4 = control/white light at (20 lux) provided on 20L:4D schedule and T5 = negative control; 12 hours dark: 12 hours day light. Cobb strain broilers were used in a Complete Randomize Design with 6 replicates. WG, water/feed intake, feed conversion ratio (FCR), mortality, behavior and welfare were assessed. At 35 d, significantly (p<0.05) highest body weight ($2,147.06g{\pm}99$) was recorded by T3. Lowest body weight ($1,640.55g{\pm}56$) and FCR (1.34) were recorded by T5. Skin weight was the only carcass parameter showed a significant (p<0.05) influence giving the highest (56.2 g) and the lowest (12.6 g) values for T5 and T1 respectively. Reduced welfare status indicated by significantly (p<0.05) higher foot pad lesions, hock burns and breast blisters was found under T3, due to reduced expression of behavior. Highest walking ($2.08%{\pm}1%$) was performed under T1 in the evening during 29 to 35 days. Highest dust bathing ($3.01%{\pm}2%$) was performed in the morning during 22 to 28 days and highest bird interaction (BI) ($4.87%{\pm}4%$) was observed in the evening by T5 during 14 to 21 days. Light $intensity{\times}day$ $session{\times}age$ interaction was significantly (p<0.05) affected walking, dust bathing and BI. Light intensity significantly (p<0.05) affected certain behaviors such as lying, eating, drinking, standing, walking, preening while lying, wing/leg stretching, sleeping, dozing, BI, vocalization, idling. In conclusion, birds essentially required provision of light in the night for better performance. Exposed to 5 lux contributed to higher WG, potentially indicating compromised welfare status. Further researches are suggested to investigate RL intensity based lighting regimen that favors for both production and welfare of Broilers.

• 미생물학회지
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• 제38권3호
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• pp.173-179
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• 2002

발광 박테리아인 Photobacterium species의 lux 오페론에서 발견된 riboflavin 생합성에 관여하는 유전자들(ribI,II,III,IV)의 기능을 조사하였다. 대장균에서 이들 유전자가 포함된 재조합 플라스미드를 발현시켰을 때 상당량의riboflavin이 합성되는 것을 확인하였으며, 또한 이들 유전자들(ribI,II,III,IV)의 기능을 riboflavin에 대하여 종속 영양체인 대장균 돌연변이주(BSV 11,18)를 이용한 유전학적인 방법과 생화학적 방법으로 분석한 결과, 이들은 각각 riboflavin synthase, 3,4-dihydroxy-2-butanone 4-phosphate (DHBP) synthase, lumazine synthase, GTP cyclohydrolase II활성도를 갖는 단백질을 코드하는 것으로 밝혀졌다. 이는Photobacterium species의 riboflavin 유전자 체계가 riboflavin 생합성에 관여하는 모든 5개의 유전자들이 한 오페론에 존재하는 Bacillus subtilis와 주요 riboflavin 유전자들이 분리되어 있는 대장균과는 다른, 중간적인 형태를 갖는다는 것을 나타낸다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권6호
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• pp.395-399
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• 2000

Five different freeze-dried recombinant bioluminescent bacteria were used for the detection of cellular stresses caused by endocrine disrupting chemicals. These strains were DPD2794 (recA::luxCDABE), which is sensitive to DNA damage, DPD2540 (fabA::luxCDABE), sensitive to cellular membrane damage, DPD2511 (katG::luxCDABE), sensitive to oxidative damage, and TV1061 (grpE::luxCDABE), sensitive to protein damage. GC2, which emits bioluminescence constitutively, was also used in this study. The toxicity of several chemicals was measured using GC2. Damage caused by known endocrine disrupting chemicals, such as nonyl phenol, bisphenol A, and styrene, was detected and classified according to toxicity mode, while others, such as phathalate and DDT, were not detected with the bacteria. These results suggest that endocrine disrupting chemicals are toxic in bacteria, and do not act via an estrogenic effect, and that toxicity monitoring and classification of some endocrine disrupting chemicals may be possible in the field using these freeze-dried recombinant bioluminescent bacteria.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.187-190
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• 2000

본 연구는 5가지의 발광성 미생물을 이용하여 유해 방사선으로 알려져 있는 ${\gamma}-rays$가 여러가지 cellular stresses 중, 특히 유전자 손상과 생물막 손상을 유발하였는데, 이들의 손상 정도가 총 방사선량과 상관관계가 있음을 발생하는 bioluminescence 로써 확인하였다. 뿐만 아니라, 선량률의 변화를 통하여 방사선으로 인한 유전자 손상 및 일반적인 독성 효과가 큰 영향을 받는 것을 확인하였는데, 선량률 증가에 따라 이들 손상정도가 증가하는 것으로 보아 선량률이 genetic 및 radioprotecion에 심각한 영향을 미치는 것을 확인하였다.

• 대한환경공학회지
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• 제34권9호
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• pp.630-636
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• 2012

본 연구는 해양미세조류 Dunaliella salina을 LED 광원을 이용하여 배양한 연구로 배양에 필요한 최적의 파장, 광도, 공기공급속도 및 배양온도를 찾기 위하여 수행되었다. 낮은 배양온도에서보다는 높은 온도에서 성장이 빨랐으나, 본 실험에서는 $22^{\circ}C$에서 가장 잘 성장하는 것으로 확인되었다. LED 파장에 따른 성장실험에서는 백색 LED가 배양에 가장 효과적이었으며, 광도에 따른 실험에서는 실험 시 고려한 3가지 광도 중(3,000, 6,300, 8,000 Lux) 3,000 Lux에서 최대 세포농도인 1.30 g/L를 나타내었다. 공기공급속도의 증가는 Dunaliella salina의 성장속도와 반비례의 관계를 나타내었는데, 공기를 공급하지 않은 경우의 비증식속도 $1.12day^{-1}$와 비교하면 공기공급은 본 미세조류의 성장에 저해요인으로 작용함을 알 수 있었다.