Bioinformatics and high throughput approach to create genomic resources for the study of bovine immunobiology
2002
Coussens, Paul M. | Nobis, William
The advent of technologies such as real-time reverse transcriptase polymerase chain reaction (RT-PCR) and cDNA microarrays herald a new era in the study of biological systems. In immunobiology, these advances have begun to impact studies of infectious diseases, inflammatory processes, and immune cell function. However, a lack of genetic reagents for domestic and companion animals has precluded widespread application of new technologies to studies in these systems. We have recently described development of cDNA microarrays for studying bovine immunobiology. Although powerful in revealing genes involved in immunological phenomena in cattle, these resources were limited by a lack of genes known to function in immune responses from other species, such as mouse and human. To address this shortcoming, we used a combination of bioinformatics and high throughput RT-PCR to create amplicons representing over 270 bovine genes whose orthologs in other species were known to function in immune responses. Amplified gene segments were prepared from cDNA representing RNA isolated from either unstimulated or concanavalin A (ConA) stimulated peripheral blood mononuclear cells (PBMCs). In total, 276 genes were amplified from cDNA representing unstimulated bovine PBMC RNA or from cDNA representing ConA stimulated bovine PBMC RNA. A web-accessible resource (http://gowhite.ans.msu.edu/public_php/gd-bovine-immunology.php) has been created to assist in dissemination of this novel resource. The web-accessible resource contains information on gene name, the forward and reverse primers used to amplify each segment, expected product size, and if the gene was found in unstimulated PBMCs or only in ConA stimulated PBMCs. Gene names appear as hyperlinks to the Genbank pages representing the bovine gene or expressed sequence tag (EST) used to generate each primer pair.
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