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Global Gene Expression Analysis Using a Zebrafish Oligonucleotide Microarray Platform
Published on: August 10, 2009
Transcriptome sequencing and development of an expression microarray platform for the domestic ferret.
Carl E Bruder1, Suxia Yao, Francis Larson
1Southern Research Institute, Birmingham, AL 35205, USA. carl.bruder@gmail.com
BMC Genomics
|April 21, 2010
Summary
Researchers developed new tools for ferret functional genomics by creating a large expressed sequence tags (EST) dataset and a ferret-specific microarray. This advances research into ferret respiratory diseases like influenza.
Area of Science:
- Genomics
- Molecular Biology
- Animal Models
Background:
- Ferrets are valuable models for respiratory diseases, including influenza.
- Limited reagents hinder molecular and immunological studies in ferrets.
- This study addresses the need for enhanced genomic resources for ferrets.
Purpose of the Study:
- To generate an extensive expressed sequence tags (EST) dataset for ferrets.
- To develop a ferret-specific microarray platform for functional genomic analysis.
- To identify potential housekeeping genes for consistent gene expression studies.
Main Methods:
- Parallel sequencing of a normalized ferret cDNA library from multiple tissues (blood, liver, lung, spleen, brain).
- Assembly of sequence reads into partial ferret genes.
- Development and application of a ferret-specific microarray.
- Validation using quantitative real-time PCR assays.
Main Results:
- Over 500,000 sequence reads generated, assembled into approximately 16,000 partial ferret genes.
- A ferret-specific microarray platform was created using assembled genes and existing GenBank sequences.
- Tissue-specific gene expression patterns were identified and validated.
- 41 ferret genes with stable transcription profiles were identified as potential housekeeping genes.
Conclusions:
- The developed tools enable functional genomic analysis in ferrets.
- This resource facilitates the creation of new reagents for the ferret model.
- Advances in ferret genomics will support research in respiratory diseases and other fields.
