4

Environmental Benefits of Molecular Biotechnology

1. 2. 3. 4.

1. 2. 3. 4.

Emailgdroam@sun.epa.gov.tw

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)

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

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)

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

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)

10

200

(SNP)

(law enforcement)

(infrastructure)(networking)

(PMNPre-Manufacture

Notice)GEMs (Sayler & Ripp, 2000)

(Environmental Science & Technology, 1999)

6.

RNA

16S rRNA

T-RFLP

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)

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

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

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?,

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.