Transgene integration into the same chromosome location can produce alleles that express at a predictable level, or alleles that are differentially silenced.
In an effort to control the variability of transgene expression in plants, we used Cre-lox mediated recombination to insert a gus reporter gene precisely and reproducibly into different target loci. Each integrant line chosen for analysis harbors a single copy of the transgene at the designated target site. At any given target site, nearly half of the insertions give a full spatial pattern of transgene expression. The absolute level of expression, however, showed target site dependency that varied up to 10-fold. This substantiates the view that the chromosome position can affect the level of gene expression. An unexpected finding was that nearly half of the insertions at any given target site failed to give a full spatial pattern of transgene expression. These partial patterns of expression appear to be attributable to gene silencing, as low gus expression correlates with DNA methylation and low transcription. The methylation is specific for the newly integrated DNA. Methylation changes are not found outside of the newly inserted DNA. Both the full and the partial expression states are meiotically heritable. The silencing of the introduced transgenes may be a stochastic event that occurs during transformation.
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Plant Gene Expression Center, U.S. Department of Agriculture-Agricultural Research Service, Albany, California 94710, USA.
Lee
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Kobayashi
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Holappa
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LD
Albert
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Ow
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DW
D000483:Alleles; D019175:DNA Methylation; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression; D020868:Gene Silencing; D017343:Genes, Plant; D017930:Genes, Reporter; D018745:Genome, Plant; D005966:Glucuronidase / Q000235:genetics; D008540:Meiosis; D008957:Models, Genetic; D016254:Mutagenesis, Insertional; D010947:Plants, Toxic; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D011995:Recombination, Genetic; D014018:Tissue Distribution; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic; D019076:Transgenes
D018744:DNA, Plant; D012333:RNA, Messenger; D018749:RNA, Plant; D005966:Glucuronidase