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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.
Potato modified to synthesise fusion spider silk fragment/ elastin protein
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7 lines: pCB301-Kan-So1-100xELP-31, -33, -37, -38, -39, -44 and -50
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- Organization: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) | BCH-CON-DE-49376-2Organization:Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)Academic or research instituteCorrensstrasse 3Gatersleben,
06466 , GermanyPhone: +49 (0)39482 5-0,Fax: +49 (0)39482 5139,Email: info@ipk-gatersleben.de,
Potatoes pCB301-Kan-SO1-100xELP produce a fusion protein consisting of the repetitive part of the spider silk protein MaSPI and an artificial elastin which is mainly observed in tubers. Female golden silk orbweaver spiders Nephila clavipes incorporate MaSpI protein within the very strong and elastic silk of the main threads of the spider net which is of high industrial interest. The artificial elastin 100xELP has high similarity to human elastin and consists of oligomeric repeats of the pentapeptide Val-Pro-Gly-Xaa-Gly (Xaa – any amino acid but proline). 100xELP exhibits a good ability to be solved in aqueous solution depending on salt concentration and temperature. It can undergo an inverse transition, i.e. the protein is soluble in an aqueous solution below its transition temperature but insoluble above this critical value. This characteristic is delegated to its fusion partner SO1 resulting in easy isolation and purification of spider silk protein from the biomass of potato tubers.
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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-12106-6 Organism Solanum tuberosum (Potato, SOLTU)Crops
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Potato modified to synthesise fusion spider silk/ elastin protein| Resistance to antibiotics (Kanamycin), Use in industrial applications
pCB301-Kan-SO1-100xELP (derivate of pCB301-Kan from pBIN19)
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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-15001-5 Neomycin Phosphotransferase II | Escherichia coli (ECOLX)Protein coding sequence | Resistance to antibiotics (Kanamycin)
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BCH-GENE-SCBD-48457-2 Synthetic spider silk protein coding sequence | Nephila clavipes (Golden silk orbweaver spider)Protein coding sequence | Use in industrial applications
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BCH-GENE-SCBD-100270-6 Nopaline Synthase Gene Promoter | Agrobacterium tumefaciens (Agrobacterium)Promoter
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BCH-GENE-SCBD-100290-6 CaMV 35S terminator | Cauliflower mosaic virus (CaMV)Terminator
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BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)Promoter
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BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)Terminator
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BCH-GENE-SCBD-103025-2 LeB4 Signal peptide | Vicia faba (Broad Bean, Tick Bean, Windsor Bean, Horse Bean, Pigeon Bean, Field Bean)Transit signal
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BCH-GENE-SCBD-103554-2 c-myc-tag | Homo sapiens (HUMAN)Transit signal
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BCH-GENE-SCBD-103553-2 Synthetic elastin-like proteinProtein coding sequence | Production of medical or pharmaceutical compounds (human or animal),Use in industrial applications
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BCH-GENE-SCBD-103023-2 KDEL ER retention signalTransit signal
The introduced recombinant DNA includes two gene cassettes.
First, the fusion protein consisting of the synthetic spider silk protein SO1 and the elastin-like protein 100xELP carries the signal peptide LeB4 for translocation to the endoplasmic reticulum (ER) at the N-terminus and the KDEL signal peptide for retention in the ER at the C-terminus. The expression of this construct is controlled by the CaMV 35S promoter and –terminator. The coding sequence SO1 represents the repetitive part of the spider silk protein MaSpI; approx. 180 bp of the MaSpI gene was removed form the 3´-end. The c-myc tag serves as immunochemical evidence of the fusion protein in Western blot analyses using antibodies.
Second, a nptII gene cassette consisting of the nos promoter, the nptII gene and nos terminator serves as a selection marker system via aminoglycosid antibiotics.
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First, the fusion protein consisting of the synthetic spider silk protein SO1 and the elastin-like protein 100xELP carries the signal peptide LeB4 for translocation to the endoplasmic reticulum (ER) at the N-terminus and the KDEL signal peptide for retention in the ER at the C-terminus. The expression of this construct is controlled by the CaMV 35S promoter and –terminator. The coding sequence SO1 represents the repetitive part of the spider silk protein MaSpI; approx. 180 bp of the MaSpI gene was removed form the 3´-end. The c-myc tag serves as immunochemical evidence of the fusion protein in Western blot analyses using antibodies.
Second, a nptII gene cassette consisting of the nos promoter, the nptII gene and nos terminator serves as a selection marker system via aminoglycosid antibiotics.
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- Research
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- Purification of recombinant proteins by fusion with thermally-responsive polypeptides. [ English ]
- Production of spider silk proteins in tobacco and potato.pdf [ English ]
- Purification of spider silk-elastin from transgenic plants and application for human chondrocyte proliferation [ English ]
- A mini binary vector series for plant transformation [ English ]
| Record type | Field | Record(s) | |
|---|---|---|---|
| Country's Decision or any other Communication | LMO identification | 1 | |
| Risk Assessment generated by a regulatory process | Living modified organism(s) | 1 | |
| Living Modified Organism | Related LMO(s) | 1 | |
