DNA insert from MON87427 PV-ZMAP1043
Transcription of 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) from Agrobacterium tumefaciens commences from the Cauliflower mosaic virus (CaMV) enhanced 35S promoter and ends at the A. tumefaciens nopaline synthase (nos) gene terminator. The transcript contains a Zea mays heat shock protein 70 (hsp70) intron, Arabidopsis thaliana N-terminal chloroplast transit peptide sequence, and cp4-epsps.  The CaMV enhanced 35S promoter-hsp70 combination promotes gene expression in female and vegetative tissues, but not in male reproductive tissues (pollen microspores and tapetum).

Note:
- Southern blot analyses indicate that a single copy of the T-DNA was inserted at a single site in the parental maize genome and no plasmid vector backbone sequences were detected to have been integrated. DNA sequencing analyses further indicated that the expected T-DNA sequences were integrated.
-The cp4-epsps coding sequence is the codon optimized coding sequence of the aroA gene from Agrobacterium sp. strain CP4 encoding CP4 EPSPS.


DNA insert from MIR162 vector pNOV1300
In the parental MIR162 maize, a variant of the native B. 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 the Z. mays ubiquitin gene promoter and then transcribes vip3Aa20 followed by intron 9 of Z. mays phosphoenolpyruvate carboxylase, before terminating at the CaMV 35S terminator. A second expression cassette, containing the E. 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 MON87411 vector PV-ZMIR10871
The MON87411 genome contains an RNA interference (RNAi) cassette targeting Diabrotica virgifera virgifera, a Bacillus thuringiensis Cry3Bb1 cassette and an Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps) cassette.

Transcription of the RNAi cassette commences from the Cauliflower mosaic virus 35S enhanced promoter and terminates at the Pisum sativum ribulose bisphosphate carboxylase small chain 2 terminator. The transcript initially contains a Zea mays heat shock protein 70 intron, which contributes to enhanced expression in vegetative tissues of the plant, and two partial coding sequences of the D. virgifera virgifera Snf7p gene, which encodes the SNF7 subunit of the ESCRT-III complex. The two Snf7p sequences are in an inverted orientation, separated by a 150-nucleotide intervening sequence, which allows base pairing between the inverted sequences and hairpin RNA formation post-transcription, which then triggers an RNAi response. Due to RNAi processing, small interfering RNA molecules (roughly 21-23 nucleotides in length) will be produced and thus no translation into protein will occur from this cassette.

Transcription of the cry3Bb1 is under control of the Z. mays physical impedance induced protein promoter and Triticum aestivum (wheat) heat shock protein 17.3 terminator. The transcript also contains a wheat 5' untranslated leader from chlorophyll a/b-binding protein and Oryza sativa actin 1 intron for enhanced expression of the transgene. Expression of cp4-epsps is under control of an O. sativa alpha tubulin promoter and terminator. The transcript additionally contains Arabidopsis thaliana chloroplast targeting peptide 2 to sequester the protein to the chloroplast.

Note:
- Sequencing, PCR and bioinformatic analyses indicate that a single, intact insertions of the three gene cassettes occurred in the parental line.
- No plasmid backbone was detected.


Kindly refer to the parental LMO records for more information.