DNA insert from MON810 (MON-ØØ81Ø-6) vector PV-ZMBK07
A partial insert containing Bacillus thuringiensis cry1Ab was inserted into the parental maize genome. Transcription is directed from the Cauliflower mosaic virus 35S enhanced promoter. The transcript contains a Zea mays heat shock protein 70 (ZmHsp70) intron and the coding sequence of cry1Ab. ZmHsp70 enhances expression of cry1Ab.

Note:
- The coding sequence of cry1Ab has been codon optimized for expression in plants. The codon optimization did not result in any changes to the amino acid sequence relative to the native sequence.
- Southern blot analysis indicated that a single partial insert is found within the parental genome.
- Southern blot analysis did not detect the presence of the Escherichia coli neomycin phosphotransferase II gene nor any DNA from plasmid PVZMGT10 (containing genes for glyphosate tolerance - cp4-epsps).
- ELISA protein analysis and feeding assays indicated expression of Cry1Ab in the parental line.


DNA insert from MIR162 (SYN-IR162-4) vector pNOV1300
In the parental MIR162 maize, a variant of the native Bacillus thuringiensis vegetative insecticidal protein 3Aa (vip3Aa20), named vip3Aa19, which has codon changes that result in a single M129I amino acid substitution was inserted into the transformation cassette. During the transformation process an additional DNA mutation resulted in a K284Q amino acid substitution. This final form was designated the name Vip3Aa20. Transcription of vip3Aa20 commences at a Z. mays ubiquitin gene promoter and then transcribes vip3Aa20 followed by intron 9 of Z. mays phosphoenolpyruvate carboxylase, before terminating at the Cauliflower mosaic virus 35S terminator. A second expression cassette, containing the Escherichia coli phosphomannose isomerase gene, was also inserted into the parental genome. The gene is under the control of another ubiquitin promoter and transcription terminates at the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator.

Note:
- Southern blot analyses demonstrated that the T-DNA insert contains: (i) single copies of a vip3Aa20 gene and a pmi gene; (ii) two copies of the maize ubiquitin promoter; (iii) one copy of the nos terminator; and (iv) no backbone sequences from transformation plasmid pNOV1300.


DNA insert from NK603 (MON-ØØ6Ø3-6) vector PV-ZMGT32
The plant expression plasmid vector, PV-ZMGT32 contains two adjacent plant gene expression cassettes each containing a single copy of Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps). In the first (5' end) expression cassette, the cp4-epsps gene is under the transcriptional regulation of an Oryza sativa actin promoter and an A. tumefaciens nopaline synthase (nos) terminator. An O. sativa actin intron is also present in the transcript for enhanced expression of the coding sequence. The second cassette consists of another cp4-epsps gene regulated by a Cauliflower mosaic virus enhanced 35S promoter (containing a duplicated enhancer region) and a nos terminator. Similarly, an intron from the maize heat shock protein 70 (ZmHsp70) was included for enhancing expression of the coding sequence. Both promoters of the gene cassettes are expected to promoter high levels of transcription.

Note:
- The parental NK603 line contained a single, intact insertion containing both cp4-epsps gene cassettes.
- Due to restriction digest prior to particle bombardment, the vector backbone, containing E. coli neomycin phosphotransferase II and origin of replication, were not incorporated into the parental genome.


Kindly refer to the parental LMO records for more information.