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THE PLANT CELL, Vol 1, Issue 7 671-680, Copyright © 1989 by American Society of Plant Biologists
Different 3[prime] End Regions Strongly Influence the Level of Gene Expression in Plant Cells
ILW. Ingelbrecht, LMF. Herman, R. A. Dekeyser, M. C. Van Montagu and A. G. Depicker
Laboratorium voor Genetica, Rijksuniversiteit Gent, B-9000 Gent, Belgium
We have investigated the functional role of a 3[prime] end region on the
expression of a reporter gene in plant cells. In stably transformed plants,
expression of the reporter gene without a plant gene 3[prime] end is
variable and depends on the fortuitous presence of polyadenylation signals
in the downstream sequences. When the reporter gene is flanked by pBR322
DNA, 3[prime]-processing and polyadenylation occurs at (a) cryptic site(s)
within these vector sequences. Using a transient gene expression system, we
present a deletion analysis of the 3[prime] end of the octopine synthase
gene showing that the most proximal polyadenylation signal per se is not
sufficient to ensure expression but that a downstream (G)T-rich sequence is
also required. Optimal expression of the fusion gene requires more than 98
base pairs and at most 142 base pairs downstream from the most distal
polyadenylation site. We analyzed the expression of chimeric genes with
3[prime] end sequences originating from different plant genes. In the
transient expression assay, all constructs direct similar neomycin
phosphotransferase II activities. However, in stably transformed tissue,
the gene constructs displayed characteristic expression levels which varied
as much as 60-fold. This result suggests a role for 3[prime] end sequences
in post-transcriptional processes such as efficiency of 3[prime]-processing
and/or mRNA stability.
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