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Living Modified Organism (LMO)
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Insect-resistant and herbicide-tolerant maize
EN
DP910521
Yes
DP-91Ø521-2
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Organization:Corteva Agriscience ()Private sector (business and industry)9330 Zionsville RoadIndianapolis, Indiana
46268, United States of AmericaPhone: +1 833-267-8382,Fax:Email: media@corteva.com,Website: https://www.corteva.com/, -
Organization:Pioneer Hi-Bred International Inc. ()Private sector (business and industry)7100 NW 62nd Avenue PO Box 1000Johnston, Iowa
50131, United States of AmericaPhone: +1 515 535-3200,Fax:Email:Website: www.pioneer.com/,
The maize (Zea mays) was modified for the resistance to lepidoptera insects and tolerance to herbicides. For resistance to lepidoptera pests, the maize expresses Bacillus thuringiensis Cry1B.34, which forms non-specific, ion conducting pores in the apical membrane of midgut epithelial cells, leading to a disruption of the midgut epithelium and eventual death. For tolerance to glufosinate, the maize expresses Streptomyces viridochromogenes phosphinothricin N-acetyltransferase, which inactivates the herbicide through acetylation. In addition, the maize also contains a gene cassette for Escherichia coli phosphomannose isomerase, which was used as a selectable marker during transformation by allowing for the maize plants to use mannose as a carbon source.
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-246-6 Organism Zea mays (Maize, Corn, MAIZE)Crops
Inbred line PH184C
EN
PHP71012; PHP79620
EN
- Biolistic / Particle gun
- Gene editing (e.g. CRISPR-Cas, etc.)
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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-100362-7 Ubiquitin gene promoter | Zea mays (Maize, Corn, MAIZE)Promoter
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loading record details...BCH-GENE-SCBD-103927-2 Ubiquitin 5' Untranslated Region | Zea mays (Maize, Corn, MAIZE)Leader
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BCH-GENE-SCBD-103627-5 Ubiquitin Intron 1 | Zea mays (Maize, Corn, MAIZE)Intron
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BCH-GENE-SCBD-15003-7 Phosphomannose Isomerase gene | Escherichia coli (ECOLX)Protein coding sequence | Mannose tolerance,Selectable marker genes and reporter genes
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BCH-GENE-SCBD-100367-4 Proteinase inhibitor II gene terminator | Solanum tuberosum (Potato, SOLTU)Terminator
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loading record details...BCH-GENE-SCBD-116046-1 19-kDa zein gene terminator | Zea mays (Maize, Corn, MAIZE)Terminator
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BCH-GENE-SCBD-100364-5 Rice actin 1 gene promoter | Oryza sativa (Rice, ORYSA)Promoter
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BCH-GENE-SCBD-100355-6 Rice actin 1, intron | Oryza sativa (Rice, ORYSA)Intron
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BCH-GENE-SCBD-15002-4 Phosphinothricin N-acetyltransferase gene | Streptomyces viridochromogenes (STRVR)Protein coding sequence | Resistance to herbicides (Glufosinate)
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BCH-GENE-SCBD-100290-6 CaMV 35S terminator | Cauliflower mosaic virus (CaMV)Terminator
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BCH-GENE-SCBD-116047-2 Ubiquitin terminator | Sorghum bicolor (Sorghum)Terminator
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BCH-GENE-SCBD-116062-1 Gamma kafarin terminator | Sorghum bicolor (Sorghum)Terminator
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loading record details...BCH-GENE-SCBD-265284-1 MMV enhancer | Mirabilis mosaic virus (MMV)Enhancer
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BCH-GENE-SCBD-265289-1 LLDV promoter | Lamium leaf distortion virus (Lamium leaf distortion-associated virus, LLDV)Promoter
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loading record details...BCH-GENE-SCBD-265290-1 Translation initiation factor 6 intron | Zea mays (Maize, Corn, MAIZE)Intron
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BCH-GENE-SCBD-265291-1 Hydroxyproline-rich glycoprotein 5' untranslated region | Zea mays (Maize, Corn, MAIZE)Enhancer
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loading record details...BCH-GENE-SCBD-265293-1 Cry1B.34 | Bacillus thuringiensis (Bt, Bacillus, BACTU)Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths), Cotton bollworm (Helicoverpa spp.), European corn borer (Ostrinia nubilalis), Fall armyworm (Spodoptera frugiperda))
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loading record details...BCH-GENE-SCBD-265292-1 Ubiquitin 1 terminator | Oryza sativa (Rice, ORYSA)Terminator
The modified maize contains three gene cassettes: Escherichia coli phosphomannose isomerase (pmi), Streptomyces viridochromogenes phosphinothricin N-acetyltransferase (pat) and Bacillus thuringiensis cry1B.34.
The pmi coding sequence is under control of a Zea mays ubiquitin (ubi) promoter and a Solanum tuberosum proteinase inhibitor II (pinII) terminator. A 5' untranslated region and intron from ubi were included to enhance expression of pmi. An additional terminator (Z. mays 19-kDa zein gene terminator) was added between the gene cassettes to prevent transcriptional interference. Due to the nature of the promoter and presence of enhancers, high levels of transcription are expected.
The pat coding sequence is under control of an Oryza sativa actin 1 promoter and Cauliflower mosaic virus 35S terminator. An intron from the actin 1 gene was also included to enhance transcription levels. An additional two terminators (Sorghum bicolor ubiquitin and gamma kafarin terminators) were added to prevent transcriptional interference between the gene cassettes.
The cry1B.34 coding sequence is under control of a Lamium leaf distortion virus promoter and O. sativa ubiquitin 1 terminator. Two Mirabilis mosaic virus enhancers and Z. mays hydroxyproline-rich glycoprotein 5' untranslated region were added adjacent to the promoter to enhance expression from this gene cassette. Due to the presence of enhancers and the constitutive nature of the promoter, high-levels of transcription are expected.
Note:
The pmi coding sequence is under control of a Zea mays ubiquitin (ubi) promoter and a Solanum tuberosum proteinase inhibitor II (pinII) terminator. A 5' untranslated region and intron from ubi were included to enhance expression of pmi. An additional terminator (Z. mays 19-kDa zein gene terminator) was added between the gene cassettes to prevent transcriptional interference. Due to the nature of the promoter and presence of enhancers, high levels of transcription are expected.
The pat coding sequence is under control of an Oryza sativa actin 1 promoter and Cauliflower mosaic virus 35S terminator. An intron from the actin 1 gene was also included to enhance transcription levels. An additional two terminators (Sorghum bicolor ubiquitin and gamma kafarin terminators) were added to prevent transcriptional interference between the gene cassettes.
The cry1B.34 coding sequence is under control of a Lamium leaf distortion virus promoter and O. sativa ubiquitin 1 terminator. Two Mirabilis mosaic virus enhancers and Z. mays hydroxyproline-rich glycoprotein 5' untranslated region were added adjacent to the promoter to enhance expression from this gene cassette. Due to the presence of enhancers and the constitutive nature of the promoter, high-levels of transcription are expected.
Note:
- The size of the Z. mays ubiquitin promoter, 5' untranslated region and intron were not available at the time of record creation.
- The pat coding sequence was optimized for expression in maize.
- Cry1B.34 protein is 1,149 amino acids in length and has a molecular weight of approximately 129 kDa.
- The maize was developed using site-specific integration. The maize was initially transformed with the following plasmids to generate the site-specific "landing pad": PHP71012, PHP70594, PHP21139 and PHP21875. CRISPR-Cas expressed from PHP70594 directs the specific insertion (via homology-directed repair) of the "landing pad" into the maize genome. Following regeneration of the maize plants, a second transformation occurred where microparticle co-bombardment was used to introduce PHP79620, PHP5096, PHP21875 and PHP73572. Transient expression of FLP recombinase replaces some "landing pad" sequences with a recombination fragment. Thus, the final modified maize contains genetic elements from PHP71012 and PHP79620. All plasmids used in the transformations are briefly described below:
- PHP71012 ("Landing pad"): Z. mays genomic recognition site; Cre recombination site (loxP); Z. mays ubiquitin 1 promoter; flippase recombination site (FRT1); neomycin phosphotransferase II; pinII terminator; flippase recombination site FRT87; genomic recognition site.
- PHP70594 (transient; not integrated into genome): cas9; guide RNA
- PHP21139 (transient; not integrated into genome): Z. mays wuschel2 (wus2)
- PHP21875 (transient; not integrated into genome): Z. mays ovule developmental protein 2 (odp2)
- PHP79620 (trait genes): FRT1; pmi; pat; loxP; cry1B.34; FRT87
- PHP5096 (transient; not integrated into genome): flippase recombinase
- PHP73572 (transient; not integrated into genome): Z. mays wus2
- PHP21875 (transient; not integrated into genome): Z. mays odp2
- Southern-by-Sequencing analysis indicated that the modified maize contains a single insert. No unexpected rearrangements, junctions, additions or plasmid backbone sequences were detected.
- Southern blot analysis indicated that the insertion was stable across five generations.
- It is recommended to refer to the attached documents in the 'Additional information' section for more information.
EN
- Feed
- Food
EN
During the characterization of DP910521 maize, proprietary maize inbred (PH184C) and F1 hybrid (PH47K2/PH184C) were used as experimental controls.
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