PREPUPAL ACTIVATION OF THE JH ESTERASE GENE

JH 酯酶基因的蛹前激活

• 批准号: 2684176
• 负责人: GRACE M JONES
• 金额: $ 17.74万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2684176
• 关键词: Culicidae; Drosophilidae; Lepidoptera; animal extract; chemical binding; complementary DNA; esterase; gel mobility shift assay; gene induction /repression; genetic library; genetic regulatory element; genetic transcription; invertebrate embryology; juvenile hormones; molecular cloning; molecular site; nucleic acid probes; nucleic acid sequence; oligonucleotides; polymerase chain reaction; reporter genes; transcription factor; transfection

The objective of the proposed grant is to use the hormonal regulation of a metamorphosis-associated protein to study the mechanisms of transcriptional regulation of genes by juvenile hormone (JH) during insect development. In Trichoplusia ni and many other insects JH esterase is strongly and positively regulated by its own hormone, JH. This system of JH induction of JH esterase is attractive for a study on how this, and perhaps other, terpenoid hormones act to regulate gene expression of a protein and how the hormone can act to regulate gene expression during development of model insects and medically important invertebrates. The specific aims and procedures of the proposed study are: 1) isolate and sequence cDNA and genomic clone(s) for JH esterases(s) in T. ni 2) define through in vivo transformation of Drosophila, or In vitro transfection assays on T. nI cell lines, specific sequences which are necessary for hormonal activation of expression of JH esterase 3) define through gel retardation, DNase I footprinting and methyl interference specific sequences or regions associated with the JH esterase gene which bind proteins following hormonal induction The results of these experiments will be new and original data on the molecular mechanisms of action of JH to stimulate gene expression. The use of a protein which becomes highly expressed during metamorphosis as the model for study assures that the findings will further our understanding of, and ability to disrupt, a fundamental and developmentally critical point of regulation of insect development. The significance and applicability of the results is ensured by the occurrence and importance of JH in a wide range of invertebrate groups.

拟议拨款的目的是使用激素 调控一种变态相关蛋白来研究其机制 保幼激素（JH）对基因转录调控的影响 昆虫发育 在粉纹夜蛾和许多其他昆虫中， 酯酶受其自身激素JH的强烈正调控。 这种JH诱导JH酯酶的系统对于研究 这种或其他萜类激素如何调节基因 蛋白质的表达以及激素如何调节基因 在模式昆虫发育过程中的表达和医学上重要的 无脊椎动物 拟议研究的具体目标和程序如下： 第一章 分离并测序JH cDNA和基因组克隆 酯酶在T. ni （二） 通过果蝇的体内转化来定义，或 体外转染T. nI细胞系，特异性 这些序列是激素激活所必需的， JH酯酶表达 第三章 通过凝胶阻滞、DNA酶I足迹和甲基 干扰特异性序列或区域， JH酯酶基因 激素诱导后结合蛋白质 这些实验的结果将是新的和原始的数据， JH刺激基因表达的分子作用机制。 使用 一种蛋白质，在变态过程中高度表达， 研究模型保证了研究结果将进一步加深我们的理解 破坏一个基本的、对发展至关重要的 昆虫发育的调节点。 的意义和 结果的适用性是由发生和重要性来保证的 JH在无脊椎动物群中的分布