Submitted to: International Symposium on Animal Genomics for Animal Health
Publication Type: Abstract Only
Publication Acceptance Date: May 1, 2007
Publication Date: October 23, 2007
Citation: Neill, J.D., Miller, L.C., Harhay, G.P., Lager, K.M. 2007. Effect of porcine reproductive and respiratory syndrome virus on porcine alveolar macrophage function as determined using serial analysis of gene expression (SAGE) [abstract]. International Symposium on Animal Genomics for Animal Health. Paper No. PA-070. p. 58. Technical Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide and causes considerable economic loss. The major target of infection is the alveolar macrophage (AM). Infection of AMs by PRRSV causes significant changes in their function by mechanisms that are not understood. We have employed SAGE to examine the global expression of genes in PRRSV-infected AMs. Total cellular RNA was prepared from in vitro cultivated AMs at 0, 6, 12, 16 and 24 hours after infection with PRRSV strain VR-2332. SAGE libraries were prepared from these RNA samples. The libraries were sequenced to obtain at least 100,000 tags from each library. These sequences were processed through sagenhaft to account for sequencing error before generating tag:count lists. These lists were deposited into a modified Identitag database for mapping to porcine genes. These lists were also used as input into the DiscoverySpace SAGE analysis application. The tag corresponding to the viral RNAs was identified and increased in number with time. Preliminary examination of the data indicates that there are changes in gene expression occurring in the PRRSV-infected AMs. Work is ongoing to further categorize these changes into specific functions/pathways and to validate the changes by real-time PCR.