Despite recent advances in chemotherapy, the human filarial infections remain important public health problems. Recently, the great advances have been made in our understanding of the repertoire of genes expressed in these parasites. However, the lack of methods of genetic manipulation and the paucity of parasite material has limited and will continue to limit studies of these organisms. Recently, it has been demonstrated that exogenous DNA sequences can be introduced into embryos of the filarial parasite Brugia malayi, employing biolistic methods and transfection with pantropic retroviral vectors. The overall goal of this proposal will be to employ these systems to overcome the limitations imposed by the lack of traditional genetics and the inability to obtain large amounts of parasite material.
The specific aims of this proposal are: 1. To employ the biolistic transient transfection system to identify cis acting factors important in controlling gene expression in B. malayi. Parameters to be explored will include identification of a minimal promoter for transient expression in B. malayi, the effect of 5' and 3' sequences important for message processing, and the effect of the addition of synthetic introns. 2. To develop pantropic retroviral vectors as a system for stable foreign gene expression in B. malayi embryos. 3. To employ pantropic retroviral vectors expressing oncogenes to create an immortalized cell line from B. malayi. 4. To determine if heterologous DNA present in transfected B. malayi embryos is stably inherited.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI048562-03
Application #
6626394
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Program Officer
Rogers, Martin J
Project Start
2001-01-15
Project End
2004-12-31
Budget Start
2003-01-01
Budget End
2003-12-31
Support Year
3
Fiscal Year
2003
Total Cost
$243,500
Indirect Cost
Name
University of Alabama Birmingham
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
063690705
City
Birmingham
State
AL
Country
United States
Zip Code
35294
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Xu, Shulin; Liu, Canhui; Tzertzinis, George et al. (2011) In vivo transfection of developmentally competent Brugia malayi infective larvae. Int J Parasitol 41:355-62
Tzertzinis, George; Egana, Ana L; Palli, Subba Reddy et al. (2010) Molecular evidence for a functional ecdysone signaling system in Brugia malayi. PLoS Negl Trop Dis 4:e625
Liu, Canhui; Oliveira, Ana; Chauhan, Chitra et al. (2010) Functional analysis of putative operons in Brugia malayi. Int J Parasitol 40:63-71
Liu, Canhui; Chauhan, Chitra; Unnasch, Thomas R (2010) The role of local secondary structure in the function of the trans-splicing motif of Brugia malayi. Mol Biochem Parasitol 169:115-9
Liu, Canhui; Chauhan, Chitra; Katholi, Charles R et al. (2009) The splice leader addition domain represents an essential conserved motif for heterologous gene expression in B. malayi. Mol Biochem Parasitol 166:15-21
de Oliveira, Ana; Katholi, Charles R; Unnasch, Thomas R (2008) Characterization of the promoter of the Brugia malayi 12kDa small subunit ribosomal protein (RPS12) gene. Int J Parasitol 38:1111-9
Ghedin, Elodie; Wang, Shiliang; Spiro, David et al. (2007) Draft genome of the filarial nematode parasite Brugia malayi. Science 317:1756-60

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