Trichoplusia ni single nucleopolyhedrovirus (TnSNPV), a 134,394-bp double-stranded DNA group II Nucleopolyhedrovirus, is pathogenic to the lepidopteran T. ni. TnSNPV transcription is temporally regulated and divided into three promoter sequence-dependent classes (early, late and very late genes). A viral oligonucleotide DNA microarray containing all potential (144) viral genes of TnSNPV was designed to investigate global viral gene expression during cell infection. Total BT1-Tn-5B1-4 cellular mRNAs extracted between 0 and 72 h posttransfection with TnSNPV genomic DNA were hybridized to the microarray. Initial average expression of early genes was detected between 12 and 24 h posttransfection while late genes were mainly detected between 24 and 72 h posttransfection. The microarray expression profiling data verified many computer predicted promoter assignments. K-means clustering was used to sort the 144 genes based on their temporal expression pattern similarities. This clustering resulted in the confirmation and temporal class assignment of previously unidentified genes and promoters.