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Modified Organism
ARSOlux Biosensor
Record information and status
Record ID
104942
Status
Published
Date of creation
2013-06-24 08:37 UTC (german_bch@bvl.bund.de)
Date of last update
2013-10-01 12:28 UTC (german_bch@bvl.bund.de)
Date of publication
2013-10-01 13:21 UTC (dina.abdelhakim@cbd.int)

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Living Modified Organism identity
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.

LMO name
ARSOlux Biosensor
Transformation event
E. coli DH5α-2697
Developer(s)
Helmholtz-Zentrum für Umweltforschung
Helmholtz Centre for Environmental Research (UFZ)
Permoserstr. 15
Leipzig, Saxony
Germany, 04318
Phone:+49 341 235-0
Fax:+49 341 235-1468
Email:info@ufz.de
Url:Helmholtz Center for Environmental Research
Description
The ARSOlux biosensor system is intended to be used as a test system to determine the concentration of arsenic compounds in waters/drinking water plants.

When these cells come in contact with arsenic compounds, the compounds enter the cells and bind to the ArsR protein. As a result, the repressor loses its ability to bind to DNA, initiating the expression of the luciferase genes. The intensity of the light emitted is proportional to the concentration of arsenic compounds and can be determined using a luminometer
Recipient Organism or Parental Organisms
The term Recipient organism refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas Parental organisms refers to those that were involved in cross breeding or cell fusion.
Escherichia coli - ECOLX
Characteristics of the transformation process
Vector
pSB403-arsR
Techniques used for the modification
  • Direct DNA transfer
Genetic elements construct
 
ArsR Promoter
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ArsR binding site
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ArsR gene
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ArsR binding site
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luxCDABE genes
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Further details
Notes regarding the genetic elements introduced or modified in this LMO
Derivative of the K12 strain with additional defects in the expression of recA and endAI. The recA mutation leads to the inability to homologously recombine DNA, resulting in the stabilisation of transferred recombinant plasmids. The endAI mutation considerably reduces endonuclease I activity, facilitating the isolation of DNA from cells.

The gene arsR including its promoter was transferred to the DH5α strain. The gene stems from the E. coli-specific plasmid R773 and encodes for a transcriptional regulator with binding sites for arsenic compounds. Furthermore, reporter genes were inserted into the recipient organisms using a vector. These are the luxCDABE genes required for the expression of luciferase from Photorhabdus luminescens.

This system employs the pSB403-arsR plasmid in the test bacteria. The vector background of this plasmid is the pRK415 plasmid, which is in turn derived from the RK2 plasmid from Klebsiella aerogenes. It contains an oriV, which enables a broad host range replication in Gram-negative bacteria, an oriT to start the plasmid transfer during a triparental conjugation and a tetracycline-resistance gene cartridge. The luxCDABE genes were inserted into pRK415 and the resulting plasmid was named pSB403. The expression of luxCDABE genes leads to emission of light.
LMO characteristics
Modified traits
  • Selectable marker genes and reporter genes
Common use(s)
  • Biosensor

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