Abstract

This renewal application represents a continuation of work on the control of gene expression in the parasitic protozoa, Leishmania enriettii. There are two major goals in our proposed project: 1) to understand the mechanism of transcriptional regulation in the parasite and define the function of DNA sequences involved in this regulation, and 2) to understand the role of RNA transsplicing in the regulation and expression of mRNA in the parasite. The genes for alpha and beta tubulin which are developmentally regulated in their expression in the parasite will be used as a model system for this study. They were cloned and characterized during the previous grant period. To accomplish these goals, it will be necessary to develop a transfection system for the functional analysis of DNA or RNA sequences by in vitro mutagenesis. A transient expression system has now been developed in our laboratory. It is the first to be described for protozoan parasites. This discovery will now be used to accomplish the aims of the grant.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI021365-04A2
Application #
3131406
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
1985-09-01
Project End
1993-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
4
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
Harvard University
Department
Type
Schools of Public Health
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Wong, A K; Chow, L M; Wirth, D F (1994) A homologous recombination strategy to analyze the vinblastine resistance property of the V-circle in Leishmania. Mol Biochem Parasitol 64:75-86
Wong, A K; Curotto de Lafaille, M A; Wirth, D F (1994) Identification of a cis-acting gene regulatory element from the lemdr1 locus of Leishmania enriettii. J Biol Chem 269:26497-502
Tobin, J F; Reiner, S L; Hatam, F et al. (1993) Transfected Leishmania expressing biologically active IFN-gamma. J Immunol 150:5059-69
Hendrickson, N; Sifri, C D; Henderson, D M et al. (1993) Molecular characterization of the ldmdr1 multidrug resistance gene from Leishmania donovani. Mol Biochem Parasitol 60:53-64
Chow, L M; Wong, A K; Ullman, B et al. (1993) Cloning and functional analysis of an extrachromosomally amplified multidrug resistance-like gene in Leishmania enriettii. Mol Biochem Parasitol 60:195-208
Tobin, J F; Wirth, D F (1993) Mutational analysis of a signal sequence required for protein secretion in Leishmania major. Mol Biochem Parasitol 62:243-9
Curotto de Lafaille, M A; Wirth, D F (1992) Creation of Null/+ mutants of the alpha-tubulin gene in Leishmania enriettii by gene cluster deletion. J Biol Chem 267:23839-46
Curotto de Lafaille, M A; Laban, A; Wirth, D F (1992) Gene expression in Leishmania: analysis of essential 5' DNA sequences. Proc Natl Acad Sci U S A 89:2703-7
Tobin, J F; Wirth, D F (1992) A sequence insertion targeting vector for Leishmania enriettii. J Biol Chem 267:4752-8
Henderson, D M; Sifri, C D; Rodgers, M et al. (1992) Multidrug resistance in Leishmania donovani is conferred by amplification of a gene homologous to the mammalian mdr1 gene. Mol Cell Biol 12:2855-65

Showing the most recent 10 out of 21 publications