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Structure and function of selectable and non-selectable transgenes in maize after introduction by particle bombardment
Authors:James C. Register III  David J. Peterson  Philip J. Bell  W. Paul Bullock  Ian J. Evans  Bronwyn Frame  Andrew J. Greenland  Nicole S. Higgs  Ian Jepson  Shuping Jiao  Carol J. Lewnau  Jeff M. Sillick  H. Martin Wilson
Affiliation:(1) Present address: ICI Seeds Research Department, 2369 330th St., 50244 Slater, IA, USA;(2) Zeneca Seeds, Jealott's Hill Research Station, Bracknell, Berkshire, UK;(3) Trait and Technology Development, Pioneer Hi-Bred International, Inc., 50131 Johnston, IA, USA
Abstract:Zea mays transformants produced by particle bombardment of embryogenic suspension culture cells of the genotype A188 × B73 and selected on kanamycin or bialaphos were characterized with respect to transgene integration, expression, and inheritance. Selection on bialaphos, mediated by thebar orpat genes, was more efficient than selection on kanamycin, mediated by thenptII gene. Most transformants contained multicopy, single locus, transgene insertion events. A transgene expression cassette was more likely to be rearranged if expression of that gene was not selected for during callus growth. Not all plants regenerated from calli representing single transformation events expressed the transgenes, and a non-selectable gene (uidA) was expressed in fewer plants than was the selectable transgene. Mendelian inheritance of transgenes consistent with transgene insertion at a single locus was observed for approximately two thirds of the transformants assessed. Transgene expression was typically, but not always, predictable in progeny plants-transgene silencing, as well as poor transgene transmission to progeny, was observed in some plant lines in which the parent plants had expressed the transgene.
Keywords:Cell biology  epigenetics  maize  transformation  transgenes
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