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GrainGenes Reference Report: PPS-106-459

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Reference
PPS-106-459
Title
Rice triosephosphate isomerase gene 5' sequence directs beta-glucuronidase activity in transgenic tobacco but requires an intron for expression in rice
Journal
Plant Physiology
Year
1994
Volume
106
Pages
459-467
Author
Xu Y
Yu H
Hall T
Abstract
In rice (Oryza sativa L), cytosolic triosephosphate isomerase (TPI) is encoded by a single gene TPI catalyzes a vital step in glycolysis, and RNA blots showed that the tpi gene is expressed in all vegetative tissues (root, culm, and leaves) and in rice suspension cells No effect of light on expression was detected, but submergence of rice seedlings resulted in elevated levels of TPI mRNA in roots and culms The 2767-bp 5' upstream sequence of the tpi gene was fused translationally with the beta-glucuronidase (gusA) gene, and the resulting construct, TPI-GUS, was found to express constitutive, high levels of GUS activity in transgenic tobacco (Nicotiana tabacum) plants However, the same construct yielded no GUS activity in stably transformed rice plants, and RNA blots showed that no GUS mRNA could be detected even though stable integration of functional copies of the construct was confirmed by Southern blot and genomic polymerase chain reaction analyses Transient assays using particle bombardment yielded high levels of GUS expression from the TPI-GUS construct in tobacco leaves, but essentially no expression in rice, barley, or maize leaves When the first intron of the tpi gene was included in the construct (TPI-int1-GUS), transient GUS activity was routinely obtained in rice leaves, revealing that the first intron of the rice tpi gene is crucial for its expression in rice TPI-int1-GUS also directed transient GUS expression in maize and barley leaves, but little or no activity was obtained from this construct in tobacco, tomato, or soybean leaves. These results with the rice tpi promoter are in accordance with mounting evidence that differences in gene expression exist. between monocots and dicots.
Keyword
[ Hide all but 1 of 13 ]
beta-glucuronidase
dicotyledons
enzyme activity
gene expression
introns
isomerases
monocotyledons
nicotiana tabacum
oryza sativa
phosphates
regulation
transgenic-plants
trioses

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