title: Activation of BmGSTd1 promoter and regulation by transcription factor Krüppel (Kr) in silkworm, Bombyx mori.
abstract: The Glutathione S-transferases (GSTs) are a large family of multifunctional enzymes, many of which play an important role in the detoxification of endogenous and exogenous toxic substances. In this research, firstly, we measured the rutin-induced transcriptional level of BmGSTd1 gene by using real-time quantitative RT-PCR method and dual spike-in strategy. The activities of the BmGSTd1 promoter in various tissues of silkworm were measured by firefly luciferase activity and normalized by the Renilla luciferase activity. Results showed that the activity of the BmGSTd1 promoter were highest in Malpighian tubule, followed by fat body, silk gland, hemocyte, epidermis, and midgut. The essential region for basal and rutin-induced transcriptional activity was -1573 to -931bp in Malpighian tubule and fat body of silkworm. Promoter truncation analysis using a dual-luciferase reporter assay in BmN cells showed that the region -1288 to -1202bp for BmGSTd1 gene was essential for basal and rutin-induced transcriptional activity. Sequence analysis of this region revealed several potential transcriptional regulatory elements such as Bcd and Kr. The mutation of core base of Kr site demonstrated that Kr functioned positively in rutin-mediated BmGSTd1 transcription.
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Guodong
authors.initials: G
authors.lastname: Zhao
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Binbin
authors.initials: B
authors.lastname: Wang
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Yunlei
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Du
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Chen
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Yaxiang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, China.
authors.forename: Weide
authors.initials: W
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China.
authors.forename: Qingyou
authors.initials: Q
authors.lastname: Xia
authors.affiliation: School of Biology and Basic Medical Sciences, Soochow University, Suzhou, Jiangsu 215123, China; National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu 215123, China. Electronic address: zgd_666@126.com.
authors.forename: Zhengguo
authors.initials: Z
authors.lastname: Wei
mesh_terms: D024506:5' Flanking Region / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D005216:Fat Body / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008317:Malpighian Tubules / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012431:Rutin / Q000494:pharmacology; D061987:Sf9 Cells; D015533:Transcriptional Activation / Q000187:drug effects
chemical_list: D019476:Insect Proteins; D007527:Isoenzymes; D051741:Kruppel-Like Transcription Factors; D012431:Rutin; D008156:Luciferases; D005982:Glutathione Transferase
keywords: BmGSTd1; Bombyx mori; Krüppel; Promoter; Rutin