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4
Environmental Benefits of Molecular Biotechnology
1. 2. 3. 4.
1. 2. 3. 4.
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5
1.
(on time/in-situ)
(denitrification, NO2N2)
PCR PrimernirK and nirS
rRNA-target probes, RT-PCR Braker et al,
1998; Wilson et al, 1998
100 -
150m (Simultaneous
nitrification and denitrification, SND)
(polyphosphate-accumulating organisms, PAOs)
(VFA)PAOs
(ployP)
(glycogen accumulating organisms,GAOs) PAOs
PAOs GAOs polyP
(Keller & Zeng, 2004)
(Giardia)(Cryptosporidium)
water blooms, toxin-producing bacteria, cyanobaceria
(Rudi et al, 1997)
16S~23SrRNA 29 DNA 9
(EPA, 2003)
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6
(PCR)18SrRNA5(genus-specific)
(primer) PCR
PCR DNA
(EPA, 2003)
SARS(How to head off the next SARS) (Anderson,
2003) 1990 (Human genome project)2003
(Human virome project)
SARS
SARS
2003
SARS
Double Agar Layer
(Brion et al, 2002)
PCR Polymerase chain reaction
DNA (denatural)
(primer)DNA (anneling)DNA polymerase
(extension) DNA polymerase
DNA DNA
DNADNA
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7
PCR DNA polymerase
PCR DNA
primer anneling PCR
PCR PCR
DNA RNA RNA
(reverse) RT-PCR(Reverse
Transcription-Polymerase Chain Reaction)
PCR (Quantitative competitive PCR)(Kim et al,
2002)Quantitative competitive PCR
(tube) PCR
PCR 2
(A)(Endogenous standards)
housekeeping gene DNA RNA
housekeeping
gene (Endogenous standards)
(B)(Externally added reference)
DNA RNA
primer
(cycle zero)
Real-time PCR QC-PCR
Real-time PCR QC-RT-PCR (Kim et al,
2002; Walsh et al, 2000; Snider et al, 2001)
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8
2.
(biosensors) CdCl2BaPs
TCE
(phenol) 3 - 4 l/ml(acetonitrile)
25 50 l/mlEPA, 2003
biosensor whole-cell biosensorDNA-binding assays (Sadik
& Witt, 1999)
3.
(Extremophiles)(thermophiles and
hyperthermophiles)(cold-loving microbes, psychrophiles)
(http://www.sciam.com/ by Michael T. Madigan and Barry L.
Marrs.)
A. FISH CLSM
(Wilderer et al, 2002)
B. r-RNA
(Witzig
et al, 2002; Kim et al, 2002)
C. r-RNA PCR-DGGEFISH
(Kurisu, 2002; Noda et al, 2002)
D. PCR
(Orsini et al, 2002)
(MGO)
GEMs GEBs (American
Chemical Society, 1999; Pieper & Reineke, 2000)
biphenyl
dioxygenases(BP Doxs)naphthalene doxsdioxin doxs in
vitro DNA-shuffling (Furukawa, 2000; Koeller & Wong, 2001)Deinococcus
radiodurans1956 Arthur W. Anderson
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9
(ionizing radiation, desiccation, UV radiation, oxidizing agents, starvation electrophilic mutagens.) 1.5 million rads
3000 DOE
(Microbial Genome Program) 1999
E. Coli
Dradiodurances
(White, 1999; Battista, 1999; Brim, 2000)
4.
(MNA)
LNAPLs DNAPLs
RT-PCR (Macdonald, 2000; Wilson et al, 1998)
(bioremediation) Hg+2
(Sayler & Ripp, 2000, Mejare & Bulow, 2001; Gadd, 2000)
DNA
mRNA RNA DNA
RNA (promoter)
mRNA
Peter C.K Lau Victor De
Lorenzo(Peter et al, 1999)
A.(starvation signals)
DNA mRNA
B.(hybrid)
C.
5.
DNA-chip, PCR, Genotoxic Effects (Blohm & Elie, 2001; Sarra, 1996)
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10
200
(SNP)
(law enforcement)
(infrastructure)(networking)
(PMNPre-Manufacture
Notice)GEMs (Sayler & Ripp, 2000)
(Environmental Science & Technology, 1999)
6.
RNA
16S rRNA
T-RFLP
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11
1.
(Convergent New Technologies)
NBIC(nano-bio-info-cogno)(EHS)
(nanotoxicology)
(ENPEngineered Nano Particle) (Mechanisms of Intake,
Uptake)(Mechanisms of Cellular, Molecular Events)
DNA
(1)
(2)
(3)
(newly generated nanoparticles)
(ENP)
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12
(1)
(2)
(3) (simple high-throughput screening
methods)(in vivo)
(regulation of cytokines and mediations, nucleus-signaling)
(structural and functional modification of proteins)
2.
green chemistry
RT-PCR
cytotoxicity, muta+, cancerogenesis
EN
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13
(BDS)DNA-shuffling (Wackett
& Bruce, 2000; Monticello, 2000)
3.
(phototrophic biofilm)
CO2
rRNA (molecular
marker) PCR 16S RNA DGGE
(Roeselers, et. al, 2004)
(horizontal
gene transfer, HGT) DNA 0~17%HGT
(mobile genetic elements,
MGEs)
MGEs
(plasmids, transposons, genomic islands,
conjugative transposons, and integrative plasmids) (Top & Springael, 2004)
DNA BTEX
(MGEs)
(Lipthay & Sprensen, 2004)
1. 1953
DNA 1990 (Human Genome
Project)2002
2.
(1)
36
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14
(2)
3000
a.
b.
c.
d.
a 500
(3)
5000
3.
American Chemical Society. 1999. Bioengineered bacteria show promise in mercury
removal. Aug. p.299. Anderson, N. Human Virome Project July 2003, The Journal of Emerging
Infectious Diseases. The China PostAug.3, 2003
Battista, J. R. et al. Why is Deinococcus radiodurans so resistant to ionizing radiation?,
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15
Trends in microbiology 362, Vol 7. No. 9. 1999. Blohm, D. H. & Guiseppi-Elie, A. 2001. New development in microarray technology.
Current Opnion in Biotechnology. Elsevier Science Ltd. P. 41-47. Brim, H. et al. Enginerrinf Deinococcus radiodurans for metal remediation in
radioactive mixed waste environments, Nature Biotechnology, Vol. 18, Jan. 2000. Brion, G. M., Meschke, J.S., Sobsey, M. D., 2002. F-speci.c RNA coliphages:
occurrence, types, and survival innatural waters. Water Res(36), 2419-2425. Braker et al. 1998. Development of PCR Primer System for Amplification of Nitrite
Reductase Genes (nirK and nirS) To Detect Denitrifying Bacteria in Environmental Samples. Applied and Environmental Microbiology. American Society for Microbiology. Oct. p. 3769 3775.
Environmental Science & Technology. 1999. Bt pollen Study Casts doubt on plant safety as EU halts approval of genetically modified corn. May. Aug. Oct. Dec.
Environmental Science & Technology. 2000. Testing Hurdles. Nov. p. 473-476. Gadd, G. M. 2000. Bioremedial potential of microbial mechanisms of metal
mobilization and immobilization. Current Opinion in Biotechnology. Elsevier Science Ltd. P. 271-279.
Keller, J. & Zeng, R.J. 2004. Biological Nitrogen and Phosphorus Removal process of amazing diversity and complexity. Eruropean Symposium on Environmental Biotechnology, ESEB 2004. Taylor & Francis Group, London. P. 9 13.
Kim, Y., Saad, M. G., Nancy, L. S., Tom, p. B. 2002. Comparison and verification of quantitative competitive reverse transcription polymerase chain reaction (QC-RT-PCR) and real time RT-PCR for avian leucosis virus subgroup J.J Virol.
Kim, S.B., Goodfellow, M., Application of Oligonucleotide Probes for the Detection of Thiothrix Spp. in Activated Sludge Plants Treating Paper and Board Mill Waster, Water Science and Technology, 2002;46:559-564.
Kim, S.B. et al. Application of oligonucleotide probes for the detection of Thiothrix spp. in activated sludge plants treating paper and board mill wastes, Water Science and Technology, Vol 46 No. 1-2, pp559-564, 2002.
Koeller, M. K. & Wong C.H. 2001. Enzymes for Chemical Synthesis. Nature. Vol. 409.
Kurukawa, K. 2000. Engineering dioxygenases for efficient degradation of environmental pollutants. Current Opinion in Biotechnology. Elsevier Science Ltd. P. 244-249.
Kreyling, W. G. et al. Health Implication of Nanoparticles, Proceedings of International Symposium on Environment Nanotechnology, Tapei, 2004.
Macdonald, J. A. 2000. Evaluating Nature Attenuation for Groundwater Cleanup. Environmental Science & Technology. American Chemical Society. Aug. P. 346-353.
Mejare, M. M. & Bulow, L. 2001. Metal-binding Proteins and peptides in Bioremediation and phytoremediation of heavy metals. Trends in Biotechnology. Vol. 19.
Monticello, D. J. 2000. Biodesulfurization and the upgrading of petroleum distillates. Current Opinion in Biotechnology. Elsevier Science Ltd. P. 540-546.
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Noda, N. et al. PCR-DGGE analysis of denitrifying bacteria in a metallurgic wastewater treatment process, Water Science and Technology, Vol 46 No.1-2, pp333-336,2002.
Oberdorster, G. Nanotoxicology: An Emerging Discipline, Proceedings of International Symposium on Environmental Technology, Tapei, 2004.
Orsini, M., Laurenti, P., Boninti, D., A Molecular Typing Approach for Evaluating Bioaerosol Exposure in Wastewater Treatment Plant Workers, Water Research, 2002; 36:1375-1378.
Peter C.K. Lau and victor De Lorengo, 1999. Genetic Engineering: The frontier of Bioremediation. EST.
Pieper, D. H. & Reineke, W. 2000. Engineering bacteria for bioremediation. Current Opinion in Biotechnology. Elsevier Science Ltd. P.262-270.
Roco, M. C. Converging Technology and Their Societal Implications, Proceedings of International Symposium on Environmental Technology, Tapei, 2004.
Rudi et al. 1997. Detection of Toxin-Producing Cyanobacteria by use of Paramagnetic Beads for Cell Concentration and DNA Purification. Applied and Environmental Microbiology. American Society for Microbiology. Jan. 1998. p. 34-37.
Sadik, O.A. & Witt, D.M. 1999. Monitoring Endocrine-Disrupting Chemicals. Environmental Science & Technology. Sept. p. 368-374.
Sayler, G. S. & Ripp, S. 2000. Field Applications of Genetically Engineered Microorganisms for Bioremediation Processes. Current Opinion in Biotechnology. Elsevier Ltd. P. 286-289.
Savva, D. Use of DNA Fingerprinting to Detect Genotoxic Effects. Ecotoxicology and Environmental Safety. Academic Press. P. 103-106.
Snider, J., V., Mark, A. W., Izidore, S. L., 2001. Human disease characterization: real-time quantitative PCR and analysis of gene expression. DDT Vol.6, No.20, 1062-1067.
Wackett, L. P. & Bruce, N. C. 2000. Environmental Biotechnology Towards Sustainability. Current Opinion in Biotechnology. Elsevier Science Ltd. P. 229-231.
Walsh, K., North, J., Barker, I., Boonham, N., 2000. Detection of different strains o Potato virus Y and their mixed infections using competitive fluorescent RT-PCR. J. Virol. Methods 91, 167-173.
White, C. et al. Genome Sequence of the Radioresistant Bacterium Deinococcus radiodurans R1, Science, Vol. 286, 19 Nov. 1999.
Wilderer, P. A., Bungartz, H.J., Lemmer h., Modern Scientific Method and Their Potential in Wastewater Science and Technology, Water Research, 2002; 36:370-393.
Witzig, R., Manz, W., Rosenberger, S., Micobiological Aspects of a Bioreactor with Submerged Membranes for Aerobic Treatment of Municipal Wastewater, Water Research, 2002;36:394-402.
91-EPA-NSC-01-B003.