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Mol Biol Rep ; 40(2): 1701-10, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23090481

RESUMO

Cytochrome P450s (CYPs) are widespread proteins that interact with exogenous chemicals from the diet or the environment. CYP9A subfamily genes are important in the silkworm Bombyx mori. We previously reported transcriptional levels of two CYP9A genes in different tissues and their responses to sodium fluoride (NaF). In this study, promoter truncation analysis using a dual-luciferase reporter assay in B. mori ovary cells (BmN) showed that the regions -1,496 to -1,102 bp for CYP9A19, and -1,630 to -1,210 bp for CYP9A22 were essential for basal transcriptional activity. Sequence analysis of these regions revealed several transcriptional regulatory elements but no typical promoter elements. Promoter activities were regulated after NaF induction and with an obvious dose effect. Although the dual-luciferase assay has been widely used to determine the activity of a given promoter in cell lines, problems with it still exist. Our results indicate that both plasmid size and construct protocols affect the experimental results.


Assuntos
Bombyx/enzimologia , Sistema Enzimático do Citocromo P-450/genética , Regulação Enzimológica da Expressão Gênica , Proteínas de Insetos/genética , Animais , Sequência de Bases , Bombyx/genética , Linhagem Celular , Clonagem Molecular , Técnicas de Silenciamento de Genes , Genes Reporter , Luciferases de Vaga-Lume/biossíntese , Luciferases de Vaga-Lume/genética , Luciferases de Renilla/biossíntese , Luciferases de Renilla/genética , Dados de Sequência Molecular , RNA Interferente Pequeno/genética , Análise de Sequência de DNA , Fluoreto de Sódio/farmacologia , Transfecção
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