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Functional analysis of cis-acting DNA elements required for expression of the SL RNA gene in the parasitic protozoan Leishmania amazonensis
Year:
1994
Source of publication :
Nucleic Acids Research
Authors :
Aly, Radi
;
.
Volume :
22
Co-Authors:
Agami, R., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Aly, R., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Halman, S., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Shapira, M., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Facilitators :
From page:
1959
To page:
1965
(
Total pages:
7
)
Abstract:
DNA sequences, that control expression of the spliced leader (SL) RNA gene in the parasitic protozoan Leishmania amazonensis, were mapped by block substitution mutagenesis. In the absence of a functional in vitro system for transcription, no promoter elements have yet been identified in this organism. We therefore developed an alternative in vivo approach, in which the SL RNA gene was tagged and then subjected to a series of linker scanning mutations. Each tagged and mutated SL RNA construct was introduced into parasite cells via the pX transfection vector, and was examined for expression of the tagged SL RNA followed by characterization of its transcriptional start site. The replacement of a critical DNA element was expected to prevent expression of the tagged SL RNA. We found that the putative SL RNA promoter is complex and includes two elements: one is located upstream to the coding region, between positions - 30 to - 70; and the other is located between -10 to +10, and includes transcribed sequences. In addition to the functional relationship between the SL RNA and vertebrate U snRNAs, we found structural similarities in their regulatory elements, which may possibly indicate a common evolutionary ancestry for these molecules. © 1994 Oxford University Press.
Note:
Related Files :
Animal
Base Sequence
DNA flanking region
gene expression
gene mapping
Molecular Sequence Data
RNA
RNA gene
Show More
Related Content
More details
DOI :
10.1093/nar/22.11.1959
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
28272
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:37
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Scientific Publication
Functional analysis of cis-acting DNA elements required for expression of the SL RNA gene in the parasitic protozoan Leishmania amazonensis
22
Agami, R., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Aly, R., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Halman, S., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Shapira, M., Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot 76100, Israel
Functional analysis of cis-acting DNA elements required for expression of the SL RNA gene in the parasitic protozoan Leishmania amazonensis
DNA sequences, that control expression of the spliced leader (SL) RNA gene in the parasitic protozoan Leishmania amazonensis, were mapped by block substitution mutagenesis. In the absence of a functional in vitro system for transcription, no promoter elements have yet been identified in this organism. We therefore developed an alternative in vivo approach, in which the SL RNA gene was tagged and then subjected to a series of linker scanning mutations. Each tagged and mutated SL RNA construct was introduced into parasite cells via the pX transfection vector, and was examined for expression of the tagged SL RNA followed by characterization of its transcriptional start site. The replacement of a critical DNA element was expected to prevent expression of the tagged SL RNA. We found that the putative SL RNA promoter is complex and includes two elements: one is located upstream to the coding region, between positions - 30 to - 70; and the other is located between -10 to +10, and includes transcribed sequences. In addition to the functional relationship between the SL RNA and vertebrate U snRNAs, we found structural similarities in their regulatory elements, which may possibly indicate a common evolutionary ancestry for these molecules. © 1994 Oxford University Press.
Scientific Publication
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