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J.Health Sci., 51(2), 197-201, 2005
Frequency of Mutations of the Transgene, which might
Result in the Loss of the Glyphosate-Tolerant Phenotype, was Lowered in Roundup Ready® Soybeans
Takeshi Ogasawara,a Yukie
Chikagawa,a Fumihiro
Arakawa,b Asami
Nozaki,a Yoshio Itoh,a Kazuo
Sasaki,c Hironori Umetsu,c Takahiro
Watanabe,d Hiroshi
Akiyama,d Tamio
Maitani,d Masatake Toyoda,e Hiroshi
Kamada,f Yukihiro
Goda,d and Yoshihiro Ozeki*,
a
aDepartment of Biotechnology, Faculty of Technology, Tokyo
University of Agriculture and Technology, 2-24-16 Naka-cho,
Koganei, Tokyo 184-8588, Japan, bSan-Ei Gen F.F.I., Inc.,
1-1-11 Sanwa-cho, Toyonaka, Osaka 561-8588, Japan,
cDepartment of Clinical Pharmacy, Faculty of Pharmaceutical
Sciences, Aomori University, 2-3-1 Kobata, Aomori 030-0943,
Japan, dNational Institute of Health Sciences, 1-18-1
Kamiyoga, Setagaya, Tokyo 158-8501, Japan,
eDepartment of Food and Health Sciences, Faculty of Human Life Sciences,
Jissen Women's University, 4-1-1 Osakaue, Hino, Tokyo
191-8510, Japan, and fInstitute of Biological Sciences, University
of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8572,
Japan
Polymerase chain reaction (PCR) primers were
prepared to amplify the DNA fragment between the genomic DNA sequence adjacent to the
5'-integration site of Roundup
Ready® (RR) soybeans neighboring the transgene and the parts of the coding region of the
transgene, together with the primer set for the
internal host gene, the alpha' subunit of
beta-conglycinin storage protein gene
(Cong gene). Using the primers for the transgene and
Cong gene, the DNA fragments were amplified from the individual genomic DNAs prepared
from 72 samples of RR soybean isolated from imported
soybean seeds labeled "not segregated." Although the
frequency of alterations of the nucleotide sequences in
both the transgene and Cong gene were almost the
same, the mutations that caused alterations to the
amino acid sequence were more highly repressed in the transgene than in the
Cong gene. In the nucleotide sequence upstream of the coding region of the
transgene, the number of alterations of the nucleotide
in the proximal promoter region was smaller than that
in the further upstream region, suggesting that the
mutants missing or being weak glyphosate-tolerance
by an alteration of the critical nucleotide sequences in
the promoter or coding region might be discarded
artificially. It is supposed that the selective bias on the
transgene might be extremely high, which indicates
that the nucleotide sequence of the transgene might
be stable and maintained in inbred RR soybean lines.
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