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Modified Organism
Barley modified for the production of LL-37 peptide
Record information and status
Record ID
115699
Status
Published
Date of creation
2020-09-01 16:26 UTC (austein.mcloughlin@cbd.int)
Date of publication
2020-09-01 16:27 UTC (austein.mcloughlin@cbd.int)

Living Modified Organism identity
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.

LMO name
Barley modified for the production of LL-37 peptide
Transformation event
UBI:LL-37
Developer(s)
Palacky University Olomouc
Institute of Molecular and Translational Medicine
Palacky University Olomouc
Olomouc
Czech Republic, 77900
Phone:+420 585 632 111
Email:reception@imtm.upol.cz
Url:The Institute of Molecular and Translational Medicine, University Palacky
Description
The barley (Hordeum vulgare) was modified for the production of human LL-37 peptide, which has known broad spectrum antimicrobial activity and acts as component of the basal immune response to infection. Barley production platforms have little phenolic compound content and have low amount of proteolytic enzymatic activity.

The line is one of three lines (see "Related LMOs") being tested for bioproduction and protein purification.  This line contains a non-specific promoter and no protein purification tag sequences. Expression of the LL-37 peptide was demonstrated to be found in all tissues of the modified plant. All lines demonstrated normal phenotypes and the LL-37 peptide was shown to be bioactive.

The modified barley additionally contains an antibiotic selection marker, Escherichia coli hygromycin phosphotransferase B, for hygromycin selection during transformation.
Recipient Organism or Parental Organisms
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.
Hordeum vulgare - Barley, HORVU
Point of collection or acquisition of the recipient organism
Hordeum vulgare cultivar Golden promise
Related LMOs
Barley modified for the production of LL-37 peptide
Palacky University Olomouc Production of medical or pharmaceutical compounds (human or animal) Protein purification Resistance to antibiotics - Hygromycin Selectable marker genes and reporter genes
Show detection method(s)
Barley modified for the production of LL-37 peptide
Palacky University Olomouc Production of medical or pharmaceutical compounds (human or animal) Resistance to antibiotics - Hygromycin Selectable marker genes and reporter genes
Show detection method(s)
Characteristics of the transformation process
Vector
pBRACT214
Techniques used for the modification
  • Agrobacterium-mediated DNA transfer
Genetic elements construct
 
CaMV 35S promoter
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Hygromycin B phosphotransferase gene
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Nopaline Synthase Gene Terminator
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Ubiquitin gene promoter
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Cytokinin dehydrogenase 1 signal peptide
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LL-37 peptide
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KDEL ER retention signal
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Nopaline Synthase Gene Terminator
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Further details
Notes regarding the genetic elements introduced or modified in this LMO
Gene cassettes
The DNA insertion contain the following two gene cassettes:
1) Escherichia coli hygromycin B phosphotransferase (hph); and
2) Homo sapiens LL-37 peptide.

Gene expression
Transcription of hph is under control of the Cauliflower mosaic virus 35S promoter and the Agrobacterium tumefaciens nopaline synthase (nos) terminator. Due to the nature of the viral promoter, transcription is expected to occur at high levels.

Transcription of the human LL-37 peptide occurs from the Zea mays ubiquitin promoter and terminates at the nos terminator. The transcript contains the following (from 5' to 3'): Z. mays cytokinin dehydrogenase 1 (ZmCKX1sp) signal peptide, LL-37 peptide and a synthetic KDEL ER retention signal. The ubiquitin promoter is non-specific and thus expression is expected in all tissues of the plant. The synthetic eukaryotic KDEL sequence facilitates the transport of LL-37 peptide to the endoplasmic reticulum (ER). Prior to export from the ER, the KDEL sequence is cleaved from the peptide. ZmCKX1sp facilitates the transit of LL-37 through the ER and excretion from the plasma membrane.

Note
- Agrobacterium tumefaciens strain AG1 was used in the transformation of barley immature zygotic embryos.
- The final LL-37 peptide is not expected to retain the signal peptides.
LMO characteristics
Modified traits
  • Production of medical or pharmaceutical compounds (human or animal)
  • Selectable marker genes and reporter genes
Common use(s)
  • Pharmaceutical
  • Research
Detection method(s)
Additional information
The LL-37 peptide can be detected in all tissues of the plant (see "Molecular Farming in Barley: Development of a Novel Production Platform to Produce Human Antimicrobial Peptide LL-37").
Additional Information
Additional Information
Cathelicidin antimicrobial peptide is the only cathelicidin protein found in humans and is location on chromosome 3p21. The sequence contains 4 exons and is translated to hCAP18, a pre-pro-protein, containing signal peptide, a conserved pro-sequence (cathelin-like domain) and a C-terminal antimicrobial peptide, LL-37. The active LL-37 peptide is produced from proteolytic cleavage from hCAP18 and its primary structure is based on 37 amino acid residues (~ 18kDa), which form an amiphiphatic alpha-helix (secondary) structure.

Records referencing this document (2)
IDDescription
2record(s) found
Modified Organism2 records