Sinorhizobium meliloti modified for the expression of a reporter gene and reduced growth | BCH-LMO-SCBD-111660 | Living Modified Organism | Biosafety Clearing-House

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Living Modified Organism (LMO)
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BCH-LMO-SCBD-111660-1   |   PDF   |   Print   |  
Decisions on the LMO Risk Assessments  
last updated: 22 Feb 2017
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Sinorhizobium meliloti modified for the expression of a reporter gene and reduced growth
EN
L1
No
The genetically modified sinorhizobia constitutively express the luciferase gene from Photinus pyralis coding for the luciferin-4-monooxygenase which catalyses a light-producing reaction and is therefore used as a reporter gene.

The luciferase expression cassette is integrated into the recA gene of the genome of S. meliloti which plays a central role in natural recombination processes, DNA repair and the control of gene expression after DNA damage. Insertion of the luciferase expression cassette, disrupts the functionality of the endogenous recA gene product.

As a result of the genetic modification S. meliloti has a prolonged generation time and a growth disadvantage.
EN
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.
The S. meliloti strain Rm2011 was used for the transformation.
EN
  • Sinorhizobium meliloti modified for the expression of a reporter gene
    | Universität Bielefeld | Selectable marker genes and reporter genes
Characteristics of the modification process
EN
  • Direct DNA transfer
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-104332-2 Firefly Luciferase | Photinus pyralis (North American firefly, Common Eastern firefly, Big Dipper firefly, PHOPY)
    Protein coding sequence | Selectable marker genes and reporter genes
  • BCH-GENE-SCBD-111595-1 Neomycin phosphotransferase II promoter | Escherichia coli (ECOLX)
    Promoter
The luciferase expression cassette was inserted into the coding sequence of the recA gene by in vivo recombination, leading to a non-functional recA gene product.

The construct did not contain its own termination sequence. It was site-specifically integrated into the S. meliloti recA Gene. The insertion site is located upstream of the endogenous recA termination region, therefore, transcription termination would be controlled by that sequence.
EN
LMO characteristics
  • BCH-GENE-SCBD-111602-1 Recombinase A gene | Sinorhizobium meliloti (RHIML)
    Protein coding sequence | Changes in physiology and/or production (Growth rate)
The insertion of the luc expression cassette into the coding sequence of the recA gene leads to a loss of function of the recA gene product.
EN
  • Research
Detection method(s)
EN
Additional Information
S. meliloti needs a host plant to grow and proliferate properly. Here, the symbiotic interaction partner alfalfa, Medicago sativa, is used as a host plant.
EN
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Country's Decision or any other Communication Living modified organism(s) 1
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