DNA microarray analysis of CfMNPV genes

Temporal expression profiles of all 146 ORFs, and their complements, of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) were determined by a modified oligonucleotide based two-channel DNA microarray. Total RNA was isolated at different times post infection from Cf203 insect cells infected with CfMNPV. The cDNA was synthesized, fluorescently labeled with Cy3, and co-hybridized to the microarray chips along with Cy5-labeled CfMNPV viral genomic DNA (vgDNA), which was used as an equimolar reference standard for each gene. From the microarray data, the temporal gene expression profiles could readily be classified into four clusters based on timing of expression. Transcription of some non-coding antisense strands of the CfMNPV genes was also detected, which provided novel insights into viral gene functions. Total RNA was isolated at different times post infection from Cf203 insect cells infected with CfMNPV. The cDNA was synthesized, fluorescently labeled with Cy3, and co-hybridized to the microarray chips along with Cy5-labeled CfMNPV viral genomic DNA (vgDNA), which was used as an equimolar reference standard for each gene.