Description of the genetic material inserted into event
Microprojectile bombardment was used to co-transform secondary
plant cell embryos with two purified linear DNA fragments: a 2924
base pair fragment (PHP19340A fragment) containing the gm-fad2-1
cassette, and the 4512 base pair fragment (PHP17752A fragment)
containing the gm-hra cassette.
The gm-fad2-1 cassette includes the promoter region from the
soybean Kunitz trypsin inhibitor gene (KTi3), a fragment of the
FAD2-1 gene that corresponds to approximately 40% of the middle
portion of the FAD2-1 gene (597 bp), and the 3' untranslated region
of the KTi3 gene (KTi3 terminator).
The gm-hra cassette contains the gm-hra gene (1971 bp), which is an
optimized form of the endogenous als gene from soybean, with
transcription regulated by the S-adenosyl-L-methionine synthetase
(SAMS) constitutive promoter (1.3 kb) from soybean and with
transcription terminated by the endogenous als gene terminator (0.6
kb) from soybean.
Sequence characterization of the inserted DNA in 305423 soybean
confirms that it contains four insertions that comprise:
* Insertion 1: one truncated PHP19340A fragment with
a truncated KTi3 terminator and intact gm-fad2-1 gene fragment and
intact KTi3 promoter, one intact PHP19340A fragment, one intact
PHP17752A fragment, one truncated PHP19340A fragment with an intact
KTi3 promoter and a truncated gm-fad2-1 gene fragment, and one
truncated PHP19340A fragment with a truncated KTi3 promoter and
truncated gm-fad2-1 gene fragment.
* Insertion 2: one truncated PHP19340A fragment with
a truncated KTi3 promoter and with intact gm-fad2-1 gene fragment
and intact KTi3 terminator.
* Insertion 3: one truncated copy of the KTi3
promoter with a nonfunctional 495 bp fragment of the plasmid
* Insertion 4: two truncated PHP19340A fragments in
an inverted repeat configuration, both with a truncated KTi3
promoter and intact gmfad2-1 gene fragment and KTi3
The genetic material that is inserted in 305423 soybean is
genetically linked and segregates following a typical pattern of
Mendelian inheritance expected for a single, genetically-linked