Suppression subtractive hybridization coupled with microarray analysis to examine differential expression of genes in virus infected cells.
High throughput detection of differential expression of genes is an efficient means of identifying genes and pathways that may play a role in biological systems under certain experimental conditions. There exist a variety of approaches that could be used to identify groups of genes that change in expression in response to a particular stimulus or environment. We here describe the application of suppression subtractive hybridization (SSH) coupled with cDNA microarray analysis for isolation and identification of chicken transcripts that change in expression on infection of host cells with a paramyxovirus. SSH was used for initial isolation of differentially expressed transcripts, a large-scale validation of which was accomplished by microarray analysis. The data reveals a large group of regulated genes constituting many biochemical pathways that could serve as targets for future investigations to explore their role in paramyxovirus pathogenesis. The detailed methods described herein could be useful and adaptable to any biological system for studying changes in gene expression.
Munir
Shirin
S
Departments of Microbiology and Veterinary Biomedical Sciences, and Biomedical Genomics Center, University of Minnesota. St. Paul, MN 55108. USA.
Singh
Sushmita
S
Kaur
Kuljeet
K
Kapur
Vivek
V