DNA insert from PV-ZMAP1043
Transcription of Agrobacterium tumefaciens
5-enolpyruvylshikimate-3-phosphate synthase (cp4 epsps)
commences from the Cauliflower mosaic virus (CaMV)
enhanced 35S promoter and terminates at the A. tumefaciens
nopaline synthase (nos) terminator. The transcript
contains a Zea mays heat shock protein 70 (hsp70)
intron, an Arabidopsis thaliana N-terminal chloroplast
transit peptide sequence for chloroplast targeting of the protein
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.
DNA insert from PV-ZMIR245
Two insecticidal protein expression cassettes were inserted into
the genome. Bacillus thuringiensis cry1A.105 expression is
under the control of the CaMV 35S enhanced promoter, which first
transcribes wheat (Triticum aestivum) 5' untranslated
region of the chlorophyll a/b-binding protein (cab) and a
rice actin 1 intron before transcribing cry1A.105.
Transcription terminates at the wheat heat shock protein 17.3
terminator. Expression of the B. thuringiensis cry2Ab2
starts at the Figwort mosaic virus (FMV) promoter, which
transcribes the Zea mays heat shock protein 70
(hsp70), then the Z. mays transit peptide and the
cry2Ab2 coding sequence, before terminating at the
- The Cry2Ab2 coding sequence was modified for optimal expression
- South blot analysis confirmed that single insertions of both
cry2Ab2 and cry1A.105, as well as no vector
backbone were present and in the parent.
- A deletion removed the duplicated enhancer elements compared to
the original CaMV e35S promoter in PV-ZMIR245.
- The selectable marker, nphII, cassette was bred out of
the parental line and became not associated with this
DNA insert from PV-ZMHT507801
The Streptomyces viridochromogenes phosphinothricin
N-acetyltransferase (pat) gene is under the control of the
Andropogon gerardii ubiquitin promoter and the
Oryza sativa alpha-amylase/trypsin inhibitor terminator.
The transcript includes the A. gerardii 5' untranslated
leader sequence and an intron from ubiquitin before (5') coding
sequence of pat.
The Stenotrophomonas maltophilia dicamba monooxygenase
(dmo) gene is under control of the Peanut chlorotic
streak caulimovirus (PC1SV) full-length transcript promoter
and the Triticum aestivum (wheat) heat shock protein 17
terminator. The transcript produced contains a wheat chlorophyll
a/b-binding 5' untranslated leader sequence (for improved gene
expression), an O. sativa actin 1 untranslated region and
intron (for improved gene expression), the untranslated and
targeting region of Petunia hybrida chloroplast transit
peptide 4 (for chloroplast targeting of the protein) and
- Originally, the plasmid vector contained two T-DNA elements that
were inserted during the initial transformation event: one
containing the dmo and pat expression cassettes,
and a second containing an Agrobacterium tumefaciens
5-enolpyruvylshikimate-3-phosphate synthase (cp4-epsps)
expression cassette. The cp4-epsps expression cassette is
regulated by the O sativa actin 1 promoter and 5′
untranslated leader, O. sativa intron, the Arabidopsis
thaliana chloroplast targeting peptide 2 targeting sequence,
and the A. tumefaciens nopaline synthase 3′
untranslated region. Subsequent traditional breeding, segregation,
selection, and screening were used to isolate those plants that
contain the dmo and pat expression cassettes
(T-DNA I) and do not contain the cp4-epsps expression
cassette (T-DNA II).
- Molecular characterization of MON87419 indicated that a single
copy of T-DNA I was integrated into the maize genome at a single
intact locus that includes all expected elements within the insert,
with the exception of incomplete Right and Left Border sequences.
These analyses also showed no PV-ZMHT507801 backbone elements or
T-DNA II sequences were present in the event.
DNA insert from PHI8999A
DNA fragment PHI8999A contains two adjacent plant gene expression
cassettes for Bacillus thuringiensis cry1F and
Streptomyces viridochromogenes pat.
Transcription of cry1F is directed by the promoter and
first exon and intron of the maize (Zea mays) ubiquitin
gene and terminates at the Agrobacterium tumefaciens ORF25
Transcription of the pat gene commences from the
Cauliflower mosaic virus (CaMV) 35S promoter and ends at
the CaMV 35S terminator.
- The coding sequence of both genes has been optimized to achieve a
high level of expression in maize.
- The sequences of the complete cry1F and pat are
identical to those in the original plasmid.
- The CRY1F protein includes the F604K (phenylalanine to lysine at
position 604) amino acid substitution, which was introduced to
create a specific restriction site for cloning purposes.
DNA insert from 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
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 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 peroxidise 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 line