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Whole Genome Sequencing and Annotation of Representatvie Strains of Spiroplasma Mirum


The overall objective of this proposal is to obtain the complete sequences of the genomes of three spiroplasma strains, two strains implicated in transmissible spongiform encephalopathies (TSE) in sheep and other ruminants, and a putative non-pathogenic strain. <P>Secondary objectives of this proposal are to 1) educate and train Louisiana undergraduate students, veterinary students, graduate students and high-school teachers in genome sequencing techniques and annotation through hands-on participation, and 2) disseminate knowledge from this project to the Mollicute research community to extend the understanding of this poorly studied group.

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NON-TECHNICAL SUMMARY: Spiroplasma mirum is a potential pathogen in man, animals and crustaceans. There is evidence that S. mirum has been associated with transmissible spongiform encephalopathies (TSE) of man and animals with significant implications to agricultural practices and human health. Also, spiroplasma are generally poorly studied at the molecular level. Therefore, genome sequencing of this group of spiroplasma will directly impact human and animal health, as well as food safety. Direct comparison of the genomes of S. mirum, an S. mirum-like spiroplasma from TSE-affected ruminant brain, and a non-pathogenic spiroplasma should support evidence for the pathogenesis of TSE in both animals and man. Comparative studies by the Bastian lab of pathogenic animal spiroplasma and those infecting plants and insects will lead to the identification of S. mirum virulence genes. This should aid in the development of preclinical detection methods and potential vaccine candidates.

<P>APPROACH: Specifically we propose to 1) construct draft genomes using 454 sequencing technology with paired-end reads, 2) construct random genomic libraries to close sequence gaps and resolve ambiguities, and 3) annotate and release the complete spiroplasma chromosome sequences. In addition, the Bastian project has a number of education and outreach activities that interweave with the research. First, a Ph.D. graduate student will be trained in microbial genomics and will gain experience as a mentor for undergraduate students and in providing microbial genomics instruction to high school teachers. Second, the project will be supporting an undergraduate student through the Howard Hughes Program and a veterinary student through the LSU/SVM Summer Scholars Program as active participants in the research and in the Spiroplasma Genome workshop during the International Organization for Mycoplasmology (IOM) International Scientific Congress in 2010. Thirdly, the Bastian lab will be conducting 3-4 workshops for high school biology teachers (12-14 teachers/session), providing training with a variety of online resources which can be accessed from the classroom.

Thune, Ronald; Bastian, Frank
Louisiana State University
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