Key Laboratory of Natural Pesticide and Chemical Biology, Ministry of Education, Laboratory of Insect Toxicology, South China Agricultural University, Guangzhou, PR China.
J Biotechnol. 2012 May 31;159(1-2):115-20. doi: 10.1016/j.jbiotec.2012.02.017. Epub 2012 Mar 7.
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.
本研究通过抑制差减杂交，揭示了小菜蛾细胞系对印楝素 A（Aza）响应的差异表达基因。在 Aza 响应的 SSH 文库中，大约 270 个序列代表了 53 个不同的已鉴定基因，这些基因编码具有各种预测功能的蛋白质，基因簇的百分比分别为 26.09%（遗传信息处理）、11.41%（细胞生长和死亡）、7.07%（代谢）、6.52%（信号转导/运输）和 2.72%（免疫）。选择 11 个与鉴定基因同源的克隆通过定量实时聚合酶链反应进行确认。在验证的 11 个克隆中，除一个脂酶的转录本外，所有转录本在 ETA 收集的 SL 细胞系中均增加，而在 ETA 收集的 SL 细胞系中，其他基因的转录本均低于 UETA。这些基因被认为与 SL 细胞系对 Aza 的反应有关。这将为揭示 Aza 作用于 SL 细胞系的分子机制提供新的线索。
