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Living Modified Organism
(LMO)
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.
Tobacco modified for antibiotic resistance and GFP expression
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Nt-pDK53
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Organization:University of Rostock ()Academic or research institute, Agrobiotechnology - Faculty of Agricultural and Environmental SciencesFaculty of Agricultural and Environmental Sciences Justus-von-Liebig-Weg 6Rostock,
18059, GermanyPhone: +49 381 498 3001,Fax:Email: dekan.auf@uni-rostock.de,
Tobacco line Nt-pDK53 contains chloroplasts that were genetically modified to express the GFP reporter gene and aadA selection marker, to evaluate the pollen-mediated transfer of transgenes located in the plastid genome in cross-breeding partners of field grown tobacco.
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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-12120-4 Organism Nicotiana tabacum (Tobacco, TOBAC )Crops
Cultivar: Petit Havana (male sterile line)
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Petunia modified for antibiotic resistance| Universität Rostock | Resistance to antibiotics (Streptomycin), Selectable marker genes and reporter genes
pDK53
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- Biolistic / Particle gun
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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-109068-1 30S ribosomal protein S16 gene terminator | Nicotiana tabacum (Tobacco, TOBAC )Terminator
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BCH-GENE-SCBD-45846-4 Green Fluorescent Protein gene | Aequorea victoria (Crystal Jellyfish, Water Jellyfish, AEQVI)Protein coding sequence | Selectable marker genes and reporter genes
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BCH-GENE-SCBD-102615-4 16S rRNA gene promoter | Nicotiana tabacum (Tobacco, TOBAC )Promoter
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BCH-GENE-SCBD-109067-1 Photosystem II protein D1 gene promoter | Nicotiana tabacum (Tobacco, TOBAC )Promoter
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BCH-GENE-SCBD-15033-8 3"(9)-O-aminoglycoside adenyltransferase | Escherichia coli (ECOLX)Protein coding sequence | Resistance to antibiotics (Streptomycin)
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BCH-GENE-SCBD-102614-3 D1 thylakoid membrane protein (psbA) gene terminator | Nicotiana tabacum (Tobacco, TOBAC )Terminator
The LMO Nt-pDK53 is a transplastomic tobacco line, therefore, chloroplast DNA was genetically modified to express the antibiotic resistance gene aadA from Escherichia coli, which is under the control of the promoter and terminator of the psbA gene, and the GFP gene from Aequoria victoria whereby expression is regulated by the 16S rRNA gene promoter, in combination with a synthetic Shine–Dalgarno sequence. The 30S ribosomal protein S16 gene terminator was used at the 3' end of the GFP coding sequence.
The transformation process was conducted in two steps:
First, the gfp reporter gene was directly linked to the aadA selectable marker gene within the pDK53 vector to ensure cointegration of the two markers into the plastid genome by homologous recombination via the flanking chloroplast sequences.
Second, young leaves from sterile tobacco plants were bombarded with plasmid-coated, 0.6-μm gold particles by using a biolistic gun.
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The transformation process was conducted in two steps:
First, the gfp reporter gene was directly linked to the aadA selectable marker gene within the pDK53 vector to ensure cointegration of the two markers into the plastid genome by homologous recombination via the flanking chloroplast sequences.
Second, young leaves from sterile tobacco plants were bombarded with plasmid-coated, 0.6-μm gold particles by using a biolistic gun.
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- Research
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