Genetically Engineered Bacteria for Genotoxicity Assessment

Biran, Alva; Pedahzur, Rami; Buchinger, Sebastian; Reifferscheid, Georg; Belkin, Shimshon

doi:10.1007/978-3-540-36253-1_6

Cited by 17 publications

(4 citation statements)

References 115 publications

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“…When the designated environmental conditions are met, the transcription of the lux operon is promoted and a light signal is produced that is proportional in intensity to the magnitude of the stimulus. Examples include bacterial reporters responsive to genotoxic agents and oxidative stress, , as well as heavy metals; benzene, toluene, ethylbenzene, and xylene (BTEX); and polycyclic aromatic hydrocarbons (PAHs). − …”

Section: Introductionmentioning

confidence: 99%

Online Monitoring of Water Toxicity by Use of Bioluminescent Reporter Bacterial Biochips

Elad

Almog²,

Yagur‐Kroll

et al. 2011

Environ. Sci. Technol.

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We describe a flow-through biosensor for online continuous water toxicity monitoring. At the heart of the device are disposable modular biochips incorporating agar-immobilized bioluminescent recombinant reporter bacteria, the responses of which are probed by single-photon avalanche diode detectors. To demonstrate the biosensor capabilities, we equipped it with biochips harboring both inducible and constitutive reporter strains and exposed it to a continuous water flow for up to 10 days. During these periods we challenged the biosensor with 2-h pulses of water spiked with model compounds representing different classes of potential water pollutants, as well as with a sample of industrial wastewater. The biosensor reporter panel detected all simulated contamination events within 0.5-2.5 h, and its response was indicative of the nature of the contaminating chemicals. We believe that a biosensor of the proposed design can be integrated into future water safety and security networks, as part of an early warning system against accidental or intentional water pollution by toxic chemicals.

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Section: Introductionmentioning

confidence: 99%

Online Monitoring of Water Toxicity by Use of Bioluminescent Reporter Bacterial Biochips

Elad

Almog²,

Yagur‐Kroll

et al. 2011

Environ. Sci. Technol.

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“…Genetically engineered microorganisms (GEMs) have been constructed for environmental applications such as, bioremediation of toxic chemicals, biological pest control, plant growth promotion (Wilson andLindow 1993 andViebahn et al, 2009) and energy production. Biran et al, (2009) were used genetically engineered bacteria for genotoxicity assessment.…”

Section: Introductionmentioning

confidence: 99%

Detection and Behavior Study of Some Genetically Engineered Microorganisms Released in Soil Using Plate Counting, Gene Transfer Assay and Specific PCR Methods

Hassan¹,

Alzohairy²

2012

Journal of Agricultural Chemistry and Biotechnology

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The objective of this study was monitoring and behavior study of genetically engineered microorganisms (GEMs) that might be accidentally or deliberately released into the environments. Three methods (plate count, gene transfer and specific PCR) were used with four genetically engineered E. coli strains (A:pJan25, B: pJan25, pGWB533 and pGWB404) harboring antibiotics resistance genes with additional gfp and gus genes as a molecular markers. The GEMs were added to soil microcosms at 10 7 cells/g soil and incubated at room temperature for 35 days. After intervals time 0, 7, 14, 21, 28 and 35 days, bacterial cells were recovered and CFU/g were estimated. The number of viable cells were decreased from 10 7 to 10 3 after 28 days with A: pJan25 and pGWB533 and reached to zero at 35 days. Strain B: pJan25 was survived up to 35 days (10 3 CFU/g), while pGWB404 was disappeared after 21 days. Comparing with sterilized soil, it was found that the viable cells were alive up to 35 days (CFU/g was 10 4).The variation of CFU and presence of a viable cells in soil microcosm may due to the effect of indigenous microbial populations and the type of strain. Specific PCR was applied on the random selected colonies at 14, 21 and 28 days only, the target genes was gfp and gus. The results showed the presence of both genes in all tested colonies. This indicated that the tested GEMs could be maintained their constructed genes at long incubation time. Horizontal gene transfer was also assayed using conjugation under laboratory and soil microcosm conditions to confirm that GEMs genes were transferred to other organisms and to monitor the persistence of GEMs genes in soil. The gene transfer was started at 14 days in sterilized soil and 21 days in soil microcosm. The conjugation frequency under laboratory and sterilized soil conditions was higher than under soil microcosm condition. The results showed that the used GEMs were able to transferred three genes to recipient cells. This indicated that these genes were plasmid harboring and it were transferred to a recipient Keywords: Genetically engineered microorganisms (GEMs), horizontal gene transfer, PCR, gfp and gus genes.

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“…, 1973 ), continuous efforts have been directed towards the development, improvement and implementation of additional bacterial‐based genotoxicity assays. One group of these assays is based on the same principle of the Ames test, in that they quantify the reversion rate from a defined mutation back to the wild‐type ( Biran et al. , 2009 ; Reifferscheid and Buchinger, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

Bacterial genotoxicity bioreporters

Biran

Yagur‐Kroll

Pedahzur

et al. 2010

Microbial Biotechnology

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SummaryEver since the introduction of the Salmonella typhimurium mammalian microsome mutagenicity assay (the ‘Ames test’) over three decades ago, there has been a constant development of additional genotoxicity assays based upon the use of genetically engineered microorganisms. Such assays rely either on reversion principles similar to those of the Ames test, or on promoter–reporter fusions that generate a quantifiable dose‐dependent signal in the presence of potential DNA damaging compounds and the induction of repair mechanisms; the latter group is the subject of the present review. Some of these assays were only briefly described in the scientific literature, whereas others have been developed all the way to commercial products. Out of these, only one, the umu‐test, has been fully validated and ISO‐ and OECD standardized. Here we review the main directions undertaken in the construction and testing of bacterial‐based genotoxicity bioassays, including the attempts to incorporate at least a partial metabolic activation capacity into the molecular design. We list the genetic modifications introduced into the tester strains, compare the performance of the different assays, and briefly describe the first attempts to incorporate such bacterial reporters into actual genotoxicity testing devices.

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Genetically Engineered Bacteria for Genotoxicity Assessment

Cited by 17 publications

References 115 publications

Online Monitoring of Water Toxicity by Use of Bioluminescent Reporter Bacterial Biochips

Online Monitoring of Water Toxicity by Use of Bioluminescent Reporter Bacterial Biochips

Detection and Behavior Study of Some Genetically Engineered Microorganisms Released in Soil Using Plate Counting, Gene Transfer Assay and Specific PCR Methods

Bacterial genotoxicity bioreporters

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