title: Expression analysis and RNA interference of BmCarE-10 gene from Bombyx mori.
abstract: Carboxylesterase (CarE) is a multifunctional superfamily, and it plays important roles in xenobiotic detoxification, pheromone degradation, neurogenesis and regulating development. In this research, firstly, we measured the rutin-induced transcriptional level of BmCarE-10 gene by using real-time quantitative RT-PCR method, and dual spike-in strategy. Several possible physiological functions were certified preliminarily by RNAi experiments in silkworm. Promoter truncation analysis using a dual-luciferase reporter assay in Bombyx mori ovary cells (BmN) showed that the region -705 to -625 for BmCarE-10 gene was essential for basal and rutin-induced transcriptional activity. Sequence analysis of this region revealed several potential transcriptional regulatory elements such as Croc and Dfd. The activities of the BmCarE-10 promoter in various tissues of silkworm were also measured by firefly luciferase activity and normalized by the Renilla luciferase activity. Results showed that the activity of the BmCarE-10 promoter were highest in the Malpighian tubule, followed by fat body, silk gland, midgut, epidermis, and hemocyte. The essential region for basal and rutin-induced transcriptional activity was also -894 to -502 in Malpighian tubule and fat body of silkworm. The potential core promoters of BmCarE-10 gene in B. mori are reported for the first time in this research. Further identification of cis- and trans-elements and their role in upregulation of BmCarE-10 gene may be useful for elucidating the contribution of CarE protein to the response mechanism to rutin.
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, No. 199 Ren'ai Road, Dushu Lake Higher Education Town, Suzhou Industrial Park, Suzhou, 215123, Jiangsu, People's Republic of China.
authors.forename: Guo-dong
authors.initials: GD
authors.lastname: Zhao
authors.forename: Ming-xia
authors.initials: MX
authors.lastname: Huang
authors.forename: Yi-ling
authors.initials: YL
authors.lastname: Zhang
authors.forename: Xue-cen
authors.initials: XC
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Du
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Chen
authors.forename: Ya-xiang
authors.initials: YX
authors.lastname: Xu
authors.forename: Wei-de
authors.initials: WD
authors.lastname: Shen
authors.forename: Zheng-guo
authors.initials: ZG
authors.lastname: Wei
mesh_terms: D000818:Animals; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D043182:Carboxylesterase / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D007814:Larva; D010053:Ovary / Q000166:cytology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012431:Rutin / Q000494:pharmacology
chemical_list: D012431:Rutin; D043182:Carboxylesterase