Spinosad represents a new class of insecticides produced by Saccharopolyspora spinosa.To understand the transcription of the spinosad biosynthetic gene cluster, two promoter detection plasmids based on different reporter genes were constructed and used to detect 9 promoters in the gene cluster.In addition, the temporal transcriptional profiles of the corresponding genes controlled by the 9 promoters, together with 4 genes outside the spinosad cluster but required for the synthesis of sugar moieties in spinosad molecule, were examined by real-time PCR.