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The Lymantria dispar multinucleocapsid nuclear polyhedrosis virus (LdMNPV) gene encoding G22 was cloned and sequenced. The G22 gene codes for a 191 amino acid protein with a predicted M r of 22000. Expression of G22 in a rabbit reticulocyte system generated a protein with an M r of 24000, in close agreement with the molecular mass predicted from the nucleotide sequence. G22 is not significantly homologous to any known protein, nor is a G22 homologue present in the Autographa californica MNPV (AcMNPV). Temporal expression studies indicated that the G22 gene was transcribed at readily detectable levels in the presence of cycloheximide. Transcripts were detected immediately after the virus adsorption period and throughout the infection cycle. The early transcriptional start sites of G22 map to a sequence that resembles a subset of RNA polymerase II promoters/start sites that are found upstream of Drosophila melanogaster developmental and retrotransposon genes which lack TATA box motifs. Several consensus late baculovirus promoter/ mRNA start site sequences (ATAAG) were identified upstream of the G22 gene start codon.