Negative effect of the 5′-untranslated leader sequence on Ac transposon promoter expression

Negative effect of the 5′-untranslated leader sequence on Ac transposon promoter expression Transposable elements are used in heterologous plant hosts to clone genes by insertional mutagenesis. The Activator (Ac) transposable element has been cloned from maize, and introduced into a variety of plants. However, differences in regulation and transposition frequency have been observed between different host plants. The cause of this variability is still unknown. To better understand the activity of the Ac element, we analyzed the Ac promoter region and its 5′-untranslated leader sequence (5′ UTL). Transient assays in tobacco NT1 suspension cells showed that the Ac promoter is a weak promoter and its activity was localized by deletion analyses. The data presented here indicate that the core of the Ac promoter is contained within 153 bp fragment upstream to transcription start sites. An important inhibitory effect (80%) due to the presence of the 5′ UTL was found on the expression of LUC reporter gene. Here we demonstrate that the presence of the 5′ UTL in the constructs reduces the expression driven by either strong or weak promoters. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Negative effect of the 5′-untranslated leader sequence on Ac transposon promoter expression

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Publisher
Springer Journals
Copyright
Copyright © 1999 by Kluwer Academic Publishers
Subject
Life Sciences; Biochemistry, general; Plant Sciences; Plant Pathology
ISSN
0167-4412
eISSN
1573-5028
D.O.I.
10.1023/A:1006288503153
Publisher site
See Article on Publisher Site

Abstract

Transposable elements are used in heterologous plant hosts to clone genes by insertional mutagenesis. The Activator (Ac) transposable element has been cloned from maize, and introduced into a variety of plants. However, differences in regulation and transposition frequency have been observed between different host plants. The cause of this variability is still unknown. To better understand the activity of the Ac element, we analyzed the Ac promoter region and its 5′-untranslated leader sequence (5′ UTL). Transient assays in tobacco NT1 suspension cells showed that the Ac promoter is a weak promoter and its activity was localized by deletion analyses. The data presented here indicate that the core of the Ac promoter is contained within 153 bp fragment upstream to transcription start sites. An important inhibitory effect (80%) due to the presence of the 5′ UTL was found on the expression of LUC reporter gene. Here we demonstrate that the presence of the 5′ UTL in the constructs reduces the expression driven by either strong or weak promoters.

Journal

Plant Molecular BiologySpringer Journals

Published: Oct 19, 2004

References

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