This is the first revision of the R21 application, AI064118-01, which is in response to PA-03-080 Biodefense and Emerging Infectious Disease Research Opportunities. Enterocytozoon bieneusi is a clinically significant human enteric pathogen associated with immunodeficiencies including AIDS, and against which therapy is not readily available. Given the technical difficulties associated with the lack of in vitro and in vivo propagation methods and limited sources of spores, research on this organism has been limited. Thus, we propose to undertake an E. bieneusi genome sequence survey project which will generate several important genome-related resources (sequence, clones, libraries), which will be available to the research community.
The specific aims are as follows: 1. Construct a small insert plasmid library from which approximately 40,000 clones will be sequenced. Sequence and assembly of reads to obtain 95-98% genome coverage, which should identify the majority of genes. A fosmid library will also be constructed and a 15-fold coverage will be performed. Master clone banks of both libraries will be created. 2. Perform functional genomic analyses and annotation of the generated sequence data. a. Using available computational tools, open reading frames, non-coding sequences, introns and regulatory elements will be identified. BLAST analyses will be performed to identify genes encoding proteins with high similarity to known proteins for assignment of function. b. A comparative analysis of the E. bieneusi genomic sequence with that of the Encephalitozoon cuniculi genome sequence will be carried out. Phylogenetic analysis of the cryptic mitochondrion and its function,and the ATP transporter genes will be undertaken. c. Establishment of a website to provide access to the sequence data and clones by the scientific community.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI064118-01A1
Application #
6947442
Study Section
AIDS-associated Opportunistic Infections and Cancer Study Section (AOIC)
Program Officer
Rogers, Martin J
Project Start
2005-06-15
Project End
2007-05-31
Budget Start
2005-06-15
Budget End
2006-05-31
Support Year
1
Fiscal Year
2005
Total Cost
$327,000
Indirect Cost
Name
Tufts University
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
039318308
City
Boston
State
MA
Country
United States
Zip Code
02111
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Widmer, Giovanni; Akiyoshi, Donna E (2010) Host-specific segregation of ribosomal nucleotide sequence diversity in the microsporidian Enterocytozoon bieneusi. Infect Genet Evol 10:122-8
Akiyoshi, Donna E; Morrison, Hilary G; Lei, Shi et al. (2009) Genomic survey of the non-cultivatable opportunistic human pathogen, Enterocytozoon bieneusi. PLoS Pathog 5:e1000261
Lee, Soo Chan; Corradi, Nicolas; Byrnes 3rd, Edmond J et al. (2008) Microsporidia evolved from ancestral sexual fungi. Curr Biol 18:1675-9
Akiyoshi, Donna E; Weiss, Louis M; Feng, Xiaochuan et al. (2007) Analysis of the beta-tubulin genes from Enterocytozoon bieneusi isolates from a human and rhesus macaque. J Eukaryot Microbiol 54:38-41
Corradi, Nicolas; Akiyoshi, Donna E; Morrison, Hilary G et al. (2007) Patterns of genome evolution among the microsporidian parasites Encephalitozoon cuniculi, Antonospora locustae and Enterocytozoon bieneusi. PLoS One 2:e1277
Feng, Xiaochuan; Akiyoshi, Donna E; Sheoran, Abhineet et al. (2006) Serial propagation of the microsporidian Enterocytozoon bieneusi of human origin in immunocompromised rodents. Infect Immun 74:4424-9