中国水产科学2011，Vol.18Issue(6)：1211-1218,8.DOI:10.3724/SP.J.1118.2011.01211
牙鲆let-7基因的克隆与表达及其靶基因预测
Cloning, expression and target gene prediction of let-7 in Paralichthys olivaceus metamorphosis
摘要
Abstract
The let-7 gene is a critical regulator of developmental timing events of the larval-to-adult transition. Here, Paralichthys olivaceus let-7 was identified by a stem-loop RT-PCR. The precursor of let-7 was cloned and was proven to be highly conserved among different species. Quantitative analysis revealed that the expression levels of let-7 were generally variably enhanced during the first five metamorphic stages and peaked during the late metamorphic stages (36 dph). At the end of P. Olivaceus metamorphosis (41 dph), expression of let-7 was reduced. To study the effects of triiodothyronine (T3) on the expression of let-7, six different concentrations (0 nmol-L-1, 50 nmol·L-1, 75 nmol·L-1, 100 nmol·L-1, 200 nmol-L1) were used to treat P. Olivaceus kidney cells in vitro. Quantitative PCR showed that let-7 levels increased after T3 treatment, especially at 75 nmol-L-1, indicating that T3 regulated the expression of let-7. Further investigation of target genes demonstrated that the 3'-untranslated regions (3'-UTRs) of 29 mRNAs were possible target sites matched with let-7. These Mrna targets played key roles in diverse biological processes: development, cell proliferation, signal transduction, virus immune response, and signaling pathways. These re suits suggested that let-7 played an essential role in regulating the developmental timing of P. Olivaceus metamorphosis.关键词
牙鲆/let-7/克隆与表达/变态/甲状腺激素/靶基因Key words
Paralichthys olivaceus/ let-7/ cloning and expression/ metamorphosis/ triiodothyronine (T3)/ target gene分类
农业科技引用本文复制引用
施志仪,吴明林,付元帅..牙鲆let-7基因的克隆与表达及其靶基因预测[J].中国水产科学,2011,18(6):1211-1218,8.基金项目
国家自然科学基金项目(31172392) （31172392）
上海重点学科项目(S30701). （S30701）
