Haemobartonella Felis (Mycoplasma Haemofelis) Molecular Diagnostics and Genomic Studies

Abstract

Little was known about the genome size or structure of the haemotrophic mycoplasmas and studies of pathogenicity, adhesion, and protein production are hampered by the inability to culture H. felis and use more common genetic tools. A renewable source of H. felis DNA in the form of a large insert library was created to study the genome and search for genes related to pathogenicity and survival. The genome size of H. felis was determined to be 1200 kb using pulsed field gel electrophoresis. A bacterial artificial chromosome (BAC) was used to create a large insert library of the H. felis genomic DNA. Using hybridization experiments and fingerprinting analysis, these BACs were ordered into a genomic encyclopedia with a restriction map. The BACs were then sub-cloned into a plasmid to generate high throughput genomic sequencing surveys (GSS) data. These GSSs were compared to the four completed sequenced mycoplasma and deposited in GenBank. This represents the first description of a physical and genomic map of an uncultivable mycoplasma.