An SSH library responsive to azadirachtin A constructed in Spodoptera litura Fabricius cell lines
The present study revealed differentially expressed genes responsive to azadirachtin A (Aza) in Spodoptera litura cell line through suppression subtractive hybridization. In the Aza-responsive SSH library, approximately 270 sequences represent 53 different identified genes encoding proteins with various predicted functions, and the percentages of the gene clusters were 26.09% (genetic information processing), 11.41% (cell growth and death), 7.07% (metabolism), 6.52% (signal transduction/transport) and 2.72% (immunity), respectively. Eleven clones homologous to identified genes were selected to be confirmed through quantitative real time polymerase chain reaction. Among the eleven clones validated, all but one transcript of lipase showed an increase in SL cell line collected from ETA, whereas the transcripts of other genes were lower in the SL cell line collected from ETA compared with that of UETA. These genes were considered to be related to the response of SL cell line to Aza. These will provide a new clue to uncover the molecular mechanisms of Aza acting on SL cell line.
► A cDNA library of azadirachtin A acting on Spodoptera litura Fabricius cell line was constructed through suppression subtractive hybridization technique. ► Fifty-three differentially expressed genes were found from the Aza responsive SSH library. ► Eleven genes were verified to be regulated by Aza in QRT-PCR. ► The median inhibition concentration of Aza after 48 h exposed to Spodoptera litura cell line was examined.
Journal: Journal of Biotechnology - Volume 159, Issues 1–2, 31 May 2012, Pages 115–120