Cloning of upstream sequences responsible for cell cycle regulation of the Nicotiana sylvestris CycB1;1 gene

Cloning of upstream sequences responsible for cell cycle regulation of the Nicotiana sylvestris... To understand the mechanisms involved in the regulation of the mitotic cyclin B Nicta; CycB1;1 expression, we have cloned the Nicotiana sylvestris cyclin gene, Nicsy; CycB1;1, whose coding sequence is homologous to that of Nicta;CycB1;1 cDNA. The structure and the function of its 5′-flanking region, 1149 bp upstream of the first start codon, was analysed. By producing stably transformed cells of a synchronized culture with the Nicsy;CycB1;1 promoter/β-glucuronidase (gus) reporter gene fusion, we demonstrate that the 1149 bp promoter fragment mediates a gus transcriptional oscillation, indistinguishable from that of endogenous Nicsy;CycB1;1 cyclin B transcripts. Transient GUS activity in BY-2 protoplasts reveals that promoter activity is considerably reduced by shortening the 5′-flanking region to 538 or 320 bp. Furthermore, the 320 bp fragment no longer mediates the observed transcriptional regulation of the 1149 bp Nicsy;CycB1;1 promoter in BY-2 protoplasts isolated from synchronized cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Molecular Biology Springer Journals

Cloning of upstream sequences responsible for cell cycle regulation of the Nicotiana sylvestris CycB1;1 gene

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Publisher
Kluwer Academic Publishers
Copyright
Copyright © 1997 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:1005837931851
Publisher site
See Article on Publisher Site

Abstract

To understand the mechanisms involved in the regulation of the mitotic cyclin B Nicta; CycB1;1 expression, we have cloned the Nicotiana sylvestris cyclin gene, Nicsy; CycB1;1, whose coding sequence is homologous to that of Nicta;CycB1;1 cDNA. The structure and the function of its 5′-flanking region, 1149 bp upstream of the first start codon, was analysed. By producing stably transformed cells of a synchronized culture with the Nicsy;CycB1;1 promoter/β-glucuronidase (gus) reporter gene fusion, we demonstrate that the 1149 bp promoter fragment mediates a gus transcriptional oscillation, indistinguishable from that of endogenous Nicsy;CycB1;1 cyclin B transcripts. Transient GUS activity in BY-2 protoplasts reveals that promoter activity is considerably reduced by shortening the 5′-flanking region to 538 or 320 bp. Furthermore, the 320 bp fragment no longer mediates the observed transcriptional regulation of the 1149 bp Nicsy;CycB1;1 promoter in BY-2 protoplasts isolated from synchronized cells.

Journal

Plant Molecular BiologySpringer Journals

Published: Sep 30, 2004

References

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