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Living Modified Organism (LMO)
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Insect-resistant cotton
EN
NIBGE‑1601
No
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- Organization: National Institute for Biotechnology and Genetic Engineering (NIBGE) | BCH-CON-SCBD-258897-1Organization:National Institute for Biotechnology and Genetic Engineering (NIBGE)Academic or research instituteP.O. Box 577, Jhang RoadFaisalabad, Punjab
44000, PakistanPhone: +92 41 920 131 620,Fax: +92 041-9201322,Email: nibge.gmotesting@gmail.com,Website: http://www.nibge.org/Default.aspx,
The cotton (Gossypium hirsutum) was modified for resistance to Lepidoptera insects through the expression of Bacillus thuringiensis crystal proteins Cry1Ac and Cry2Ab2. Upon protease cleavage and activation in the insect's midgut, the bioactive core toxins form pores, which disrupt osmotic balance and eventually result in cell lysis and insect death. In addition to the insecticidal proteins, the cotton expresses an Escherichia coli neomycin phosphotransferase II cassette for kanamycin selection during transformation.
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-12080-6 Organism Gossypium hirsutum (Cotton)Crops
Coker 312 cultivar
EN
pGA482
EN
- 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-101416-6 Ti plasmid right border repeat | Agrobacterium tumefaciens (Agrobacterium)Plasmid vector
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BCH-GENE-SCBD-100270-6 Nopaline Synthase Gene Promoter | Agrobacterium tumefaciens (Agrobacterium)Promoter
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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-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)Terminator
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BCH-GENE-SCBD-101507-5 FMV 34S promoter | Figwort mosaic virus (Figwort mottle virus, FMV, CMoVb)Promoter
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BCH-GENE-SCBD-14988-7 Cry2Ab2 | Bacillus thuringiensis (Bt, Bacillus, BACTU)Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
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BCH-GENE-SCBD-103067-9 Transcript 7 gene 3' untranslated region | Agrobacterium tumefaciens (Agrobacterium)Terminator
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BCH-GENE-SCBD-100290-6 CaMV 35S terminator | Cauliflower mosaic virus (CaMV)Terminator
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BCH-GENE-SCBD-14986-6 Cry1Ac | Bacillus thuringiensis (Bt, Bacillus, BACTU)Protein coding sequence | Resistance to diseases and pests (Insects, Lepidoptera (butterflies and moths))
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BCH-GENE-SCBD-100366-6 CaMV Enhanced 35S promoter | Cauliflower mosaic virus (CaMV)Promoter
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BCH-GENE-SCBD-101415-9 Ti plasmid left border repeat | Agrobacterium tumefaciens (Agrobacterium)Plasmid vector
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BCH-GENE-SCBD-100365-6 Chloroplast transit peptide 2 | Arabidopsis thaliana (Thale cress, Mouse-ear cress, Arabidopsis, ARATH)Transit signal
The modified cotton contains three gene cassettes: Escherichia coli neomycin phosphotransferase II (nptII); Bacillus thuringiensis cry2Ab2; and B. thuringiensis cry1Ab.
The nptII coding sequence is under control of an Agrobacterium nopaline synthase promoter and terminator.
The cry2Ab2 coding sequence is under control of a Figwort mosaic virus 34S promoter and an A. tumefaciens transcript 7 gene 3' untranslated region. An Arabidopsis thaliana 5-enolpyruvylshikimate-3-phosphate synthase signal sequence (chloroplast transit peptide 2) was added to target the translated Cry2Ab2 protein to the chloroplast.
The cry1Ac coding sequence is under control of a Cauliflower mosaic virus 35S enhanced promoter and 35S terminator. Due to the duplicated enhancer regions and the constitutive nature of the promoter high levels of expression are expected.
Note:
The nptII coding sequence is under control of an Agrobacterium nopaline synthase promoter and terminator.
The cry2Ab2 coding sequence is under control of a Figwort mosaic virus 34S promoter and an A. tumefaciens transcript 7 gene 3' untranslated region. An Arabidopsis thaliana 5-enolpyruvylshikimate-3-phosphate synthase signal sequence (chloroplast transit peptide 2) was added to target the translated Cry2Ab2 protein to the chloroplast.
The cry1Ac coding sequence is under control of a Cauliflower mosaic virus 35S enhanced promoter and 35S terminator. Due to the duplicated enhancer regions and the constitutive nature of the promoter high levels of expression are expected.
Note:
- The nptII cassette was originally sourced from pGA482. The base pair sizes of the genetic elements in this cassette were not found.
EN
- Fiber/textile
- [PDF] Development of event‑specific detection method for identification of insect resistant NIBGE‑1601 cotton harboring double gene Cry1Ac‑Cry2Ab construct.PDF [ English ]
- [HTML] Development of event‑specific detection method for identification of insect resistant NIBGE‑1601 cotton harboring double gene Cry1Ac‑Cry2Ab construct [ English ]
EN
Please note that the GenBank sequence contains an additional cassette (epsps) that is not present in the NIBGE-1601 cotton line.
- Development of event‑specifc detection method for identifcation of insect resistant NIBGE‑1601 cotton harboring double gene Cry1Ac‑Cry2Ab construct.pdf [ English ]
- Development and evaluation of double gene transgenic cotton lines expressing Cry toxins for protection against chewing insect pests.pdf [ English ]
- GenBank - Synthetic construct EPSPS, Cry2Ab, and Cry1Ac genes, complete cds [ English ]
