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Notification report


General information

Notification Number
B/CZ/18/01

Member State to which the notification was sent
Czech Republic

Date of acknowledgement from the Member State Competent Authority
12/10/2018

Title of the Project
Optimization of purification of LL-37 peptide produced by
transgenic barley


Proposed period of release:
01/01/2019 to 31/12/2029

Name of the Institute(s) or Company(ies)
Usovsko a.s., ;


3. Is the same GMPt release planned elsewhere in the Community?
No

Has the same GMPt been notified elsewhere by the same notifier?
No

Genetically modified plant

Complete name of the recipient or parental plant(s)
Common NameFamily NameGenusSpeciesSubspeciesCultivar/breeding line
barleypoaceaehordeumhordeum vulgarevulgareGolden promise

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
Modification UBI:LL-37
contains peptide LL-37, hpt, UBI promoter and NOS terminator, KDEL, ZmCKX1
N-terminal signal peptide
Modification bHOR:LL-37
contains LL-37, hpt, KDEL, ZmCKX1sp, NOS, B1 hordein promoter (bHOR)
Modification bHOR:MBP_LL-37
contains LL-37, hpt, KDEL, ZmCKX1sp, bHOR, NOS, MBP and 6xHis


Genetic modification

3. Type of genetic modification:
Insertion;

In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:
Modification UBI:LL-37
Peptide LL-37 - human gene coding Cathelicidin, commercially synthesized.
Hpt (YP_006952299), the hygromycin phosphotransferase gene derived from
Escherichia coli. Source - binary vectors pBRACT209 and pBRACT214
UBI promoter (Zea mays L.) and NOS terminator (Agrobacterium tumefaciens) - to
drive over-expression of LL-37 in barley. Source - binary vector pBRACT214.
KDEL - eukaryotic signal peptide, commercially synthesized.
ZmCKX1sp - the N-terminal signal of peptide cytokinin dehydrogenase 1 from Zea
mays L., responsible for the translocation of the peptide through the
endoplasmic reticulum. Source - commercially synthesized.
Modification bHOR:LL-37
It contains, as in previous modifications, LL-37, hpt, KDEL, ZmCKX1sp and NOS.
B1 hordein promoter (bHOR) - endosperm specific promoter from barley, used for
expression of LL-37 in barley endosperm. Source - commercially synthesized.
Modification bHOR:MBP_LL-37
It contains, as in previous modifications, LL-37, hpt, KDEL, ZmCKX1sp, bHOR and
NOS.
MBP - protein expression tag for purification of LL-37 peptide. Source -
commercially synthesized.
6xHis - A polyhistidine-tag is for the purification of LL-37 peptide. Source -
commercially synthesized.


6. Brief description of the method used for the genetic modification:
Transformation of barley immature grains (Golden promise) by
Agrobacterium tumefaciens strain AGL1.


7. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination:
NA

Experimental Release

1. Purpose of the release:
The purpose is to study the stable production of LL-37 peptide in field conditions, assess the yield of the recombinant product, and to optimize the purification of this peptide from mature transgenic barley grains and also from immature grains (in milk-wax maturity).

2. Geographical location of the site:
Cadastral teritory Mohelnice, plot 2502/4, region Olomouc

3. Size of the site (m2):
14.919 m2, the area of GM plants will be much smaller, the first year less than 400 m2

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:
NA

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
The transgenic plants producing the LL-37 peptide (T1-T3 generation) were
grown in a closed phytotron and in greenhouses of the Palacký University
(CRH, Department of Molecular Biology) in Olomouc within contained GMO use management. No selective advantages or disadvantages, nor any interactions with
control plants or other organisms were observed for the selected modifications (UBI:LL-37, bHOR:LL-37 and bHOR:MBPLL-37).
The modified spring barley does not pose any risks to the environment, nor to any risks to human health or animal health


Brief description of any measures taken for the management of risks:
The work on the experimental field and all manipulation with GM plant
material will be done by trained workers of Usovsko a.s. All handling of GM
plant material will be monitored and recorded. The personnel will be
instructed about the emergency plan and the operating rules of the
experimental field prior to sowing.
The transgenic plants will be surrounded by 3 m wide border of non-modified barley. After the harvest of the GM barley, all plants including the border rows will be chopped and crushed on the site. After a proper period for germination of any seeds, the plot will be treated by a herbicide.
The minimal isolation distance from any commercially grown barley will be 100 m.
The monitoring plan will be implemented during and after the experiments, as
described in the plan. The monitoring will take place on the
experimental plot and in its immediate vicinity. In case of an accident, the monitoring will also be carried out at the site of the accident and in its immediate vicinity.


Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
Does not apply.

Final report
-

European Commission administrative information

Consent given by the Member State Competent Authority:
Not known