Deliberate release into the E.U.
environment of GMOs for any other purposes than placing on the
market:
Evaluation of transgenic potato as
bioreactor for production of recombinant spider silk under field
conditions - Institut für Pflanzengenetik und
Kulturpflanzenforschung Gatersleben |
Date of publication: February 4,
2005
Source:
http://gmoinfo.jrc.it/gmp_browse_geninf.asp
Notification
report
General information
Notification Number: B/DE/04/160
Member State: Germany
Date of Acknowledgement: 13/10/2004
Title of the Project:
Evaluation of transgenic potato as bioreactor for production
of recombinant spider silk under field conditions
Proposed period of release From:01/04/2005
To:31/10/2005
Name of the Institute(s) or Company(ies): Institut für
Pflanzengenetik und Kulturpflanzenforschung Gatersleben (IPK);
3. Is the same GMPt release planned elsewhere in the
Community?
No
4 - Has the same GMPt been notified elsewhere by the same
notifier?
No
Genetically
modified plant
1. Complete name of the
recipient or parental plant(s)
|
Common Name
|
Family Name
|
Genus |
Species
|
Subspecies
|
Cultivar/breeding line
|
| potato
|
solanaceae |
solanum |
solanum tuberosum |
tuberosum |
Albatros |
2. Description of the traits and characteristics which have
been introduced or modified, including marker genes and previous
modifications:
For the deliberate release potato plants (Solanum tuberosum),
variety Albatros, were modified by genetic engineering. The
plants were transformed by Agrobacterium-mediated gene transfer.
The transferred genes of interest are coding for spider silk
proteins and elastin-like proteins (MaSpI-100ELP, MaSpII-100ELP
and SO1-100xELP). Transgenic potato plants as bioreactors for
the production of spider silk proteins under field conditions
will be evaluated.
MaSpI and MaSpII are spider silk proteins from Nephila clavipes.
SO1 is a synthetic spider silk protein and mimics the repetitive
part of MaSpI from Nephila clavipes. 100xELP is a synthetic gene
and is homologous to human elastin. MaSpI-ELP, MaSpII-ELP and
SO1-ELP are fused at the N-terminus to the Vicia faba legumin A
signal peptide sequence. This causes the translation at the ER
associated ribosomes and the transport into the Endoplasmic
Reticulum. Between the sequences coding for MaSpI, MaSpII and
SO1, respectively, and the sequences coding for 100xELP a
sequence coding for the human c-myc-tag was inserted. The
c-myc-tag is a short amino acid sequence out of the human C-MYC
protein. It is used for the immunochemical detection of the
fusion proteins and has no enzymatic activity. The sequences
coding for the fusion proteins are C-terminally fused to the
sequence coding for the ER retention signal KDEL. This causes
retention of the recombinant proteins in the ER. The expression
of the fusion proteins is regulated by the CaMV35S promoter, an
ubiquitous promoter. This promoter is regulates the expression
of 35S transcripts of Cauliflower Mosaic Virus. For termination
of transcription the CaMV35S untranslated region at the 3énd is
used.
To differentiate between transformed and untransformed cells a
gene coding for kanamycin resistance was introduced into the
genome of the transgenic potato plants. The resistance gene is
regulated by the nopaline synthase promoter and terminator from
Agrobacterium tumefaciens. The Kanamycin resistance gene codes
for a well characterized enzyme, the neomycin
phosphotransferase. There is no hint for toxicity. Several
transgenic plants containing the neophosphotransferase coding
gene were evaluated by the US Food and Drug Adsministration
(FDA) and by the US Environmental Protection Agency and
characterized as completely save. Those plants are generally
integrated into the US agriculture. Some lines contain the
nptIII gene, that could be expressed only in bacteria, but not
in plants. The nptIII gene causes resistance against Kanamycin,
Neomycin and Amikacin. Amikacin is an important antibiotic of
value in therapy. Taking into account, that (i) the probability
of horizontal gene transfer from plants to bacteria is very low
and (ii) there is no selection pressure at the area of
deliberate release it could be assumed, that the frequence of
nptIII caused resistance in microorganisms will not be assumed
by the deliberate release.
Genetic
modification
3. Type of genetic
modification:
Insertion;
4. In case of insertion of genetic material, give the source
and intended function of each constituent fragment of the region
to be inserted:
Description of composition and origin of the transgene:
-CaMV35S promoter from Cauliflower Mosaic Virus: promoter for
expression of the gene of interest; LeB4-signal peptide from
Vicia faba L.: signal peptide for transport of transgenic
protein of interest into the Endoplasmatic Reticulum; spider
silk gene MaSpI from Nephila clavipes: structure-protein of
spider silk; spider silk gene MaSpII from Nephila clavipes:
structure-protein of spider silk; synthetic spider silk gene SO1
with homology to MaSpI from Nephila clavipes: structure-protein
of spider silk; c-myc-tag from Homo sapiens: Immunochemical
detection with antibodies in western blot, without enzymatic
activity and without DNA-adsorbtion activity; synthetic elastin
100xELP with homology to human elastin: structure-protein of
connective tissue; ER-Retionssignal KDEL from Homo sapiens:
retention of heavy chain binding protein out of Endoplasmic
Reticulum; polyadenylation signal from Cauliflower Mosaic Virus:
termination of transcription of gene of interest.
Description of composition and origin of the T-DNA: Nos promoter
from Agrobacterium tumefaciens: promoter for expression of nptII
gene; nptII-gene from E. coli: gene coding for the Neomycin
phosphotransferase; part of Ocd-gene from Agrobacterium
tumefaciens: non- functional part of the gene coding for the
Ornithin cyclodeaminase; Nos terminator: termination of
transcription; part of geneIII from phage M13: non-functional
part of the gene; part of lacZ-gene from E. coli: non-functional
part of the gene
The following parts outside of the T-DNA are also transferred:
OriV from E. coli: not functional in plants; nptII-gene from
Streptococcus faecalis: not functional in plants; trfA-gene for
replication insite of E. coli and Agrobacterium tumefaciens: not
functional in plants.
6. Brief description of the method used for the genetic
modification:
The genetic modification was brought about by
Agrobacterium-mediated gene transfer.
7. If the recipient or parental plant is a forest tree
species, describe ways and extent of dissemination and specific
factors affecting dissemination:
not applicable
Experimental
Release
1. Purpose of the release:
The purpose of the release is to evaluate potatos as
bioreactors for the production of spider silk proteins.
2. Geographical location of the site:
The field release will be at IPK Gatersleben, Saxony-Anhalt
3. Size of the site (m2):
2000
4. Relevant data regarding previous releases carried out with
the same GM-plant, if any, specifically related to the potential
environmental and human health impacts from the release:
No data regarding previous releases related to potential
environmental and human health impacts.
Environmental
Impact and Risk Management
Summary of the potential
environmental impact from the release of the GMPts:
Due to the absence of sexual compatible wild species and in
the light of the modification carried out, no obvious
environmental risks can be seen.
Brief description of any measures taken for the management of
risks:
The release site will be monitored during and after the
release. Potato lants not intended to be used for experimental
purposes will not be grown in the vivinity of the release site.
Summary of foreseen field trial studies focused to gain new
data on environmental and human health impact from the release:
not applicable |
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