Details

Autor(en) / Beteiligte

• Zhao, 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

Titel

Expression analysis and RNA interference of BmCarE-10 gene from Bombyx mori

Ist Teil von

• Molecular biology reports, 2014-03, Vol.41 (3), p.1607-1616

Ort / Verlag

Dordrecht: Springer Netherlands

Erscheinungsjahr

2014

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.