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Vol. 95, Issue 21, 12106-12110, October 13, 1998
Department of Agronomy and Plant Genetics, University of Minnesota,
411 Borlaug Hall, 1991 Upper Buford Circle, St. Paul, MN 55108
Communicated by Ronald L. Phillips, University of Minnesota, St.
Paul, MN, August 14, 1998 (received for review May 18, 1998)
Integration of transgenic DNA into the plant genome was
investigated in 13 transgenic oat (Avena sativa L.)
lines produced using microprojectile bombardment with one or two
cotransformed plasmids. In all transformation events, the transgenic
DNA integrated into the plant genome consisted of intact transgene
copies that were accompanied by multiple, rearranged, and/or
truncated transgene fragments. All fragments of transgenic DNA
cosegregated, indicating that they were integrated at single gene loci.
Analysis of the structure of the transgenic loci indicated that the
transgenic DNA was interspersed by the host genomic DNA. The number of
insertions of transgenic DNA within the transgene loci varied from 2 to
12 among the 13 lines. Restriction endonucleases that do not cleave the
introduced plasmids produced restriction fragments ranging from 3.6 to
about 60 kb in length hybridizing to a probe comprising the introduced
plasmids. Although the size of the interspersing host DNA within the
transgene locus is unknown, the sizes of the transgene-hybridizing
restriction fragments indicated that the entire transgene locus must be
at least from 35-280 kb. The observation that all transgenic lines
analyzed exhibited genomic interspersion of multiple clustered
transgenes suggests a predominating integration mechanism. We propose
that transgene integration at multiple clustered DNA replication forks
could account for the observed interspersion of transgenic DNA with
host genomic DNA within transgenic loci.
Copyright © 1998 by The National Academy of Sciences 0027-8424/98/9512106-5$2.00/0
Agricultural Sciences
Transgenic DNA integrated into the oat genome is frequently
interspersed by host DNA
*
To whom reprint requests should be addressed. e-mail:
somers{at}biosci.cbs.umn.edu.
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