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).
- 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.
- The expression of cp4-epsps from the MON87411 parental
genome is expected to overcome the tissue specific expression from
the MON87427 genome.
DNA insert from MON87411 vector PV-ZMIR10871
The MON87411 genome contains three cassettes: an RNA interference
(RNAi) cassette targeting Diabrotica virgifera virgifera,
Bacillus thuringiensis cry3Bb1 and Agrobacterium
tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase
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
- 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.
DNA insert from 59122 vector PHP17662:
Transcription of Bacillus thuringiensis cry34Ab1 starts at
Zea mays ubiquitin gene promoter and terminates at the
Solanum tuberosum proteinase inhibitor II gene terminator.
Transcription of B. thuringiensis cry35Ab1 commences from
the (Triticum aestivum (wheat) peroxidase gene promoter
and stops at another S. tuberosum proteinase inhibitor II
- The coding sequence of cry34Ab1 and
cry35Ab1 has been adapted to the codon usage in
maize as to achieve optimal expression in planta.
- The cry34Ab1 and cry35Ab1 were cloned from
B. thuringiensis strain PS149B1.
- Sequence analysis of 59122 done by the European Food Safety
Authority indicated that this LMO contains one complete copy of the
T-DNA of PHP17662 without internal rearrangements. All three gene
cassettes, cry34Ab1, cry35Ab1 and pat, are intact within the
transgenic event. The DNA sequences of the genes in 59122 are
identical to those in the original plasmid except for two
nucleotide differences in the wheat peroxidase promoter. At the 5'
T-DNA end a deletion of 22 bp is observed and at the 3' T-DNA end a
deletion of 25 bp is observed. The absence of vector backbone in
maize 59122 was also demonstrated.
For more information, kindly refer to the parental LMO