Functional genomics as a tool to study pathogenic spirochetes
| Authors | |
|---|---|
| Year of publication | 2003 |
| Type | Article in Proceedings |
| Conference | Abstrakta XVII Biological Days - Memory in living systems |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | functional genomics; pathogenic spirochetes |
| Description | Methods of functional genomics are discussed as experimental tools to study unculturable bacterial pathogens including Treponema pallidum, the causative agent of syphilis. Functional screening of a large insert T. pallidum DNA library in E. coli using a BAC cloning system can be used for screening of gene functions governed by heterologous (T. pallidum) chromosomal DNA. T. pallidum DNA-microarray- and real-time quantitative PCR-based transcriptome analyses, utilizing organisms extracted from rabbit testicular tissue and skin lesions during infection and from bacteria isolated from time-limited in vitro tissue cultures, appear to be powerful techniques in the study of gene expression levels. Results from these studies may be useful in identifying factors that limit the multiplication of T. pallidum in vitro and thus could lead to the improvement of in vitro culture techniques. Cloning and expression of the T. pallidum proteome permits a systematic evaluation of antigenicity of T. pallidum proteins. Moreover, binding interactions using two-hybrid approaches can be examined to generate large-scale protein-protein interaction maps. Whole-genome strain comparisons performed on DNA-microarrays and XL-PCR-based genome fingerprints are used to reveal genetic differences resulting in different clinical manifestations of treponemal diseases. The genome of closely related nonpathogenic T. paraluiscuniculi is used to identify genes (missing in T. paraluiscuniculi but present in T. pallidum genome) involved in the human infections. These genes are likely to code for important virulence factors and constitute promising targets for prevention/therapy. |
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