Journal article
5′ sequences essential for trans-splicing of msp (gp63) RNAs in Leishmania chagasi
Molecular and biochemical parasitology, Vol.77(1), pp.65-76
1996
DOI: 10.1016/0166-6851(96)02581-9
PMID: 8784773
Abstract
The 5′ sequences essential for spliced leader (SL) addition to RNA encoding the abundant
Leishmania surface protease gp63 were determined. A DNA segment found upstream of all
Leishmania chagasi gp63 genes was cloned in front of a luciferase gene, and luciferase activity was measured in transiently transfected
L. chagasi promastigotes. Two hundred and twenty bp of the upstream region was needed for optimal luciferase activity. Deletions and point mutations were placed in this segment to determine which nucleotides participate in the splicing events. A region containing 87% pyrimidines located between 31 and 69 bp upstream of the splice acceptor dinucleotide and containing three potential branch point A residues was highly beneficial for reported gene activity. In contrast, a 30-nt pyrimidine-rich sequence immediately upstream of the splice acceptor site did not by itself enhance luciferase activity. Luciferase activity was associated with the presence of
trans-spliced luciferase mRNA. The results suggest that sequences 31–69 bp upstream of gp63 genes enhance gene expression through provision of signals for efficient
trans-splicing.
Details
- Title: Subtitle
- 5′ sequences essential for trans-splicing of msp (gp63) RNAs in Leishmania chagasi
- Creators
- Ramesh Ramamoorthy - Department of Biochemistry, University of Iowa, Veterans' Affairs Medical Center, and Howard Hughes Medical Institute, Iowa City, IA 52242, USAJohn E Donelson - Department of Biochemistry, University of Iowa, Veterans' Affairs Medical Center, and Howard Hughes Medical Institute, Iowa City, IA 52242, USAMary E Wilson - Department of Internal Medicine and Microbiology, University of Iowa, Veterans' Affairs Medical Center, and Howard Hughes Medical Institute, Iowa City, IA 52242, USA
- Resource Type
- Journal article
- Publication Details
- Molecular and biochemical parasitology, Vol.77(1), pp.65-76
- Publisher
- Elsevier B.V
- DOI
- 10.1016/0166-6851(96)02581-9
- PMID
- 8784773
- ISSN
- 0166-6851
- eISSN
- 1872-9428
- Language
- English
- Date published
- 1996
- Academic Unit
- Microbiology and Immunology; International Programs; Epidemiology; Biochemistry and Molecular Biology; Internal Medicine
- Record Identifier
- 9984001113302771
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