| Title: | Expression analysis and RNA interference of BmCarE-10 gene from Bombyx mori |
| Author(s): | Zhao GD; Huang MX; Zhang YL; Wang XC; Du J; Li B; Chen YH; Xu YX; Shen WD; Wei ZG; |
| Address: | "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" |
| DOI: | 10.1007/s11033-013-3007-3 |
| ISSN/ISBN: | 1573-4978 (Electronic) 0301-4851 (Linking) |
| 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" |
| Keywords: | "Animals Bombyx/*genetics/metabolism Carboxylesterase/*biosynthesis/genetics Cloning, Molecular Female Gene Expression Regulation, Enzymologic/drug effects/genetics Larva Ovary/cytology/*metabolism Promoter Regions, Genetic *RNA Interference Rutin/pharmaco;" |
| Notes: | "MedlineZhao, Guo-dong Huang, Ming-xia Zhang, Yi-ling Wang, Xue-cen Du, Jie Li, Bing Chen, Yu-hua Xu, Ya-xiang Shen, Wei-de Wei, Zheng-guo eng Research Support, Non-U.S. Gov't Netherlands 2014/01/15 Mol Biol Rep. 2014 Mar; 41(3):1607-16. doi: 10.1007/s11033-013-3007-3. Epub 2014 Jan 11" |
