Project description:The domestic ferret (Mustela putorius furo) has been used as animal model for decades, largely because its susceptibility to infection with a large number of pathogens such as influenza virus, SARS Corona virus and Canine distemper virus. Despite its importance for biomedical research, little is known about the genome of the M. Furo. The number of reagents for molecular and immunological analysis is thus restricted. To circumvent this, we present here a parallel sequencing effort to produce an extensive EST dataset derived from a normalized ferret cDNA library made from mRNA from ferret blood, liver, lung, spleen and brain. We produced more than 500000 sequence reads that were assembled into over 15000 partial ferret transcripts. These ESTs were combined with the available ferret sequences in the GenBank to develop a ferret specific microarray platform. Using this array, we detected tissue specific expression patterns which were confirmed by quantitative real time PCR assays and comparison to orthologous transcription profiles of mouse and human. We also present a set of 41 ferret transcript with even transcription profile across the tested tissues, indicating their usefulness as housekeeping genes. This study paves way for development of additional reagents for analysis of the ferret model.
Project description:The domestic ferret (Mustela putorius furo) has been used as animal model for decades, largely because its susceptibility to infection with a large number of pathogens such as influenza virus, SARS Corona virus and Canine distemper virus. Despite its importance for biomedical research, little is known about the genome of the M. Furo. The number of reagents for molecular and immunological analysis is thus restricted. To circumvent this, we present here a parallel sequencing effort to produce an extensive EST dataset derived from a normalized ferret cDNA library made from mRNA from ferret blood, liver, lung, spleen and brain. We produced more than 500000 sequence reads that were assembled into over 15000 partial ferret transcripts. These ESTs were combined with the available ferret sequences in the GenBank to develop a ferret specific microarray platform. Using this array, we detected tissue specific expression patterns which were confirmed by quantitative real time PCR assays and comparison to orthologous transcription profiles of mouse and human. We also present a set of 41 ferret transcript with even transcription profile across the tested tissues, indicating their usefulness as housekeeping genes. This study paves way for development of additional reagents for analysis of the ferret model. Three biological replicates of blood, lung, spleen, liver and brain was hybridized to the ferret specific microarray.
