Knockdown of ecdysis-triggering hormone gene with a binary UAS/GAL4 RNA interference system leads to lethal ecdysis deficiency in silkworm  

作　　者：Hongjiu Dai Li Ma Jue Wang Rongjing Jiang Zhugang Wang Jian Fei 

机构地区：[1]Shanghai Research Center for Model Organism, Shanghai 201203, China [2]Institute of Plant Physiology and Ecology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China [3]Tongji University, Shanghai 200092, China

出　　处：《Acta Biochimica et Biophysica Sinica》2008年第9期790-795,共6页生物化学与生物物理学报（英文版）

摘　　要：Ecdysis-triggering hormone （ETH） is an integration factor in the ecdysis process of most insects, including Bombyx mor/（silkworm）. To understand the function of the ETH gene in silkworm, we developed an effective approach to knockdown the expression of ETH in vivo based on RNA interference （RNAi） and a binary UAS/GAL4 expression system that has been successfully used in other insect species. Two kinds of transgenic silkworm were established with this method： the effector strain with the ETH RNAi sequence under the control of UAS and the activator strain with the GAL4 coding sequence under the control of Bombyx mori cytoplasmic actin3. By crossing the two strains, double-positive transgenic silkworm was obtained, and their ETH expression was found to be dramatically lower than that of each single positive transgenic parent. Severe ecdysis deficiency proved lethal to the double-positive transgenic silkworm at the stage of pharate second instar larvae, while the single positive transgenic or wild-type silkworm had normal ecdysis. This UAS/GAL4 RNAi approach provides a way to study the function of endogenous silkworm genes at different development stages.Ecdysis-triggering hormone （ETH） is an integration factor in the ecdysis process of most insects, including Bombyx mor/（silkworm）. To understand the function of the ETH gene in silkworm, we developed an effective approach to knockdown the expression of ETH in vivo based on RNA interference （RNAi） and a binary UAS/GAL4 expression system that has been successfully used in other insect species. Two kinds of transgenic silkworm were established with this method： the effector strain with the ETH RNAi sequence under the control of UAS and the activator strain with the GAL4 coding sequence under the control of Bombyx mori cytoplasmic actin3. By crossing the two strains, double-positive transgenic silkworm was obtained, and their ETH expression was found to be dramatically lower than that of each single positive transgenic parent. Severe ecdysis deficiency proved lethal to the double-positive transgenic silkworm at the stage of pharate second instar larvae, while the single positive transgenic or wild-type silkworm had normal ecdysis. This UAS/GAL4 RNAi approach provides a way to study the function of endogenous silkworm genes at different development stages.

关 键 词：ecdysis-triggering hormone UAS/GAL4 system RNAi transgenic silkworm 

分 类 号：Q6[生物学—生物物理学]