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Living Modified Organism
(LMO)
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Hybrid aspen modified for observing horizontal gene transfer into ectomycorrhizal fungi
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Independent transformation events: T89-35S-Bar1, T89-35S-Bar3, T89-35S-Bar7, T89-35S-Bar12 and T89-35S-Bar14
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Person:BFHInstitute of Wood Research Leuschnerstraße 91 21031 Hamburg-Bergedorf Institute of Forest Genetics Sieker Landstraße 2 22927 Großhansdorf Johann Heinrich von Thünen Institute Federal Research Institute for Rural Areas, Forestry and Fisheries Bundesallee 50Brunschweig,
38116, GermanyPhone: +49 531 596 1003,Fax: + 49 531 596 1099,Email: info@thuenen.de,Related OrganizationFederal Research Centre for Forestry and Forest Products (BFH)Academic or research instituteInstitute of Wood Research Leuschnerstraße 91 21031 Hamburg-Bergedorf Institute of Forest Genetics Sieker Landstraße 2 22927 Großhansdorf Johann Heinrich von Thünen Institute Federal Research Institute for Rural Areas, Forestry and Fisheries Bundesallee 50Brunschweig,
38116, GermanyPhone: +49 531 596 1003,Fax: + 49 531 596 1099,Email: info@thuenen.de,
The bar gene from Streptomyces hygroscopicus codes for a phosphinothricin acetyltransferase (PAT) which converts L-Phosphinothricin, the active component of the herbicidal agent glufosinate-ammonium, into the derivative N-acetyl-phosphinothricin which has no herbicidal effect.
In this trial the bar gene is to be used as a marker for evidence of horizontal gene transfer from GM hybrid aspen to ectomycorrhizal fungi. This does not require the development of herbicide tolerance in the hybrid aspen. For this reason, a construct was developed which expresses the native bar gene under the control of two different promoters. A fungal promoter is located at the 5’ end of the bar gene. This is meant to allow for the selection of transgenic ectomycorrhizal fungi resulting from horizontal gene transfer. The 35S promoter of the cauliflower mosaic virus (CaMV) is positioned at the 3’ end of the bar gene in an antisense orientation. This approach is meant to prevent the development of herbicide tolerance in the GM hybrid aspen in the event that the fungal promoter is also active in plants.
Therefore, as a result of the genetic modification, herbicide resistance of the hybrid aspen is not expected. After a successful horizontal gene transfer into ectomycorrhizal fungi, however, the bar gene should be expressed and the PAT enzyme should be formed, resulting in herbicide tolerant ectomycorrhizal fungi.
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In this trial the bar gene is to be used as a marker for evidence of horizontal gene transfer from GM hybrid aspen to ectomycorrhizal fungi. This does not require the development of herbicide tolerance in the hybrid aspen. For this reason, a construct was developed which expresses the native bar gene under the control of two different promoters. A fungal promoter is located at the 5’ end of the bar gene. This is meant to allow for the selection of transgenic ectomycorrhizal fungi resulting from horizontal gene transfer. The 35S promoter of the cauliflower mosaic virus (CaMV) is positioned at the 3’ end of the bar gene in an antisense orientation. This approach is meant to prevent the development of herbicide tolerance in the GM hybrid aspen in the event that the fungal promoter is also active in plants.
Therefore, as a result of the genetic modification, herbicide resistance of the hybrid aspen is not expected. After a successful horizontal gene transfer into ectomycorrhizal fungi, however, the bar gene should be expressed and the PAT enzyme should be formed, resulting in herbicide tolerant ectomycorrhizal fungi.
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.
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BCH-ORGA-SCBD-105731-2 Organism Populus tremula x Populus tremuloides (Hybrid aspen)Trees
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pBI121-Bar
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- Agrobacterium-mediated DNA transfer
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Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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BCH-GENE-SCBD-110854-2 Glyceraldehyde 3-phosphate dehydrogenase promoter | Cochliobolus heterostrophus (Southern corn leaf blight fungus)Promoter
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BCH-GENE-SCBD-14972-12 Phosphinothricin N-acetyltransferase gene | Streptomyces hygroscopicus (STRHY)Protein coding sequence | Resistance to herbicides (Glufosinate)
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BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)Promoter
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BCH-GENE-SCBD-100270-6 Nopaline Synthase Gene Promoter | Agrobacterium tumefaciens (Agrobacterium)Promoter
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BCH-GENE-SCBD-15001-5 Neomycin Phosphotransferase II | Escherichia coli (ECOLX)Protein coding sequence | Resistance to antibiotics (Kanamycin)
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BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)Terminator
The bar gene from Streptomyces hygroscopicus coding for a phosphinothricin-acetyltransferaseis expressed under the control of two differnt promoters: the Glycerinaldehyd-3-phosphat-Dehydrogenase (GPD) promoter from Cochliobolus heterostrophus is located at the 5´ end of the gene for a transcription of the bar gene in sense orientation, the 35S promoter of the Cauliflower Mosaic Virus (CaMV) is located at the 3´ end of the gene for a transcription of the bar gene in antisense orientation.
The nptII gene coding for the neomycin phosphotransferase of the Tn5 transposon is used as a selection marker. It is expressed under the control of the promotor and the termination signal of the nopaline synthase gene (nos) Agrobacterium tumefaciens.
The transferred DNA is integrated into the genome of the recipient organism.
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The nptII gene coding for the neomycin phosphotransferase of the Tn5 transposon is used as a selection marker. It is expressed under the control of the promotor and the termination signal of the nopaline synthase gene (nos) Agrobacterium tumefaciens.
The transferred DNA is integrated into the genome of the recipient organism.
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- Research
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