SAGE tag based cDNA microarray analysis during larval to pupal development and isolation of novel cDNAs in Bombyx...

Many genes act together during the complex process of insect larval and pupal development. 20-hydroxyecdysone interacts with juvenile hormone to control insect growth and development and then activates several transcription factors, i.e., Broad, E74, and E75, and, subsequently, the late target genes. To investigate this phenomenon, we used serial analysis of gene expression (SAGE) tag-based cDNA microarray analysis to monitor the global gene expression profile during larval development and larva-pupa metamorphosis of the silkworm Bombyx mori. Of the 330 clones that were dotted to the chip, 267 were obtained by generating longer cDNA fragments from SAGE tags for gene identification, and the others were obtained from SAGE tag-matched genes or expressed sequence tags from public databases. According to the gene expression profile, the genes were classified into 12 clusters using a self-organizing map analysis. The results were partially confirmed using real-time reverse transcription-polymerase chain reaction. We obtained 22 full-length cDNAs using rapid amplification of 5' cDNA ends, of which eight genes were novel in the silkworm. Our results indicated that use of a cDNA microarray based on SAGE tags is effective for identifying and examining some low-expression genes associated with insect development.