Legionella pneumophila is an intracellular bacterium that causes an acute form of pneumonia called Legionnaires' disease. Segregation analyses using macrophages from susceptible and resistant inbred mice previously indicated that a single genetic locus, named Lgn1 , could determine permissiveness to intracellular replication of L. pneumophila. A positional cloning strategy was undertaken, which makes use of genetic and molecular biology techniques to identify the gene responsible for a particular phenotype, based mostly on its location within a chromosome. The work described in this thesis covers three aspects of Lgn1: (1) Building upon the work of others, the Lgn1 genetic interval was narrowed to 0.32 cM within distal mouse chromosome 13. The corresponding 140 Kb Lgn1 physical interval contains only two known transcripts: the Neuronal Apoptosis Inhibitor Protein ( Naip) genes Naip2 and Naip5. (2) The expression profile of the Lgn1 candidates was investigated both at the mRNA and protein levels. Expression of both Naip2 and Naip5 in mouse macrophages strengthened their candidacy for the Lgn1 locus. (3) Transfer of BAC clones from the critical interval into transgenic mice was successfully used to functionally complement the Lgn1 susceptibility phenotype of A/J mice with cloned DNA from non-permissive 129X1 or C57BL/6J origins. Two independent rescuing BAC clones were identified, with a 56-Kb overlap where the entire Lgn1 transcript must lie. The only known full-length transcript coded in this reduced genomic region is Naip5. / Thus, in our last publication we have proposed that Naip5 (recently named Birc1e) is the gene within the Lgn1 locus responsible for differential permissiveness to intracellular L. pneumophila replication in mice.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.85151 |
| Date | January 2004 |
| Creators | Diez, Eduardo |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Biochemistry.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 002198661, proquestno: AAINR12831, Theses scanned by UMI/ProQuest. |
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