PREPUPAL ACTIVATION OF THE JH ESTERASE GENE

拟议拨款的目标是使用荷尔蒙 一种变形相关蛋白的调控及其机制的研究 保幼激素(JH)对基因转录调控的研究 昆虫发育。在粉纹夜蛾和许多其他昆虫JH 酯酶受其自身激素JH的强烈正向调节。 这种JH诱导JH酯酶的体系对于研究JH酯酶具有重要意义 这种，也许还有其他的萜类激素是如何作用于调控基因的？ 蛋白质的表达以及激素如何作用于基因调控 模式昆虫发育过程中的表达及其医学意义 无脊椎动物。 拟议研究的具体目标和程序如下： 1)分离并测序JH的cDNA和基因组克隆(S) 毛滴虫中的酯酶(S) 2)通过体内转化果蝇，或在 T.ni细胞系的体外转染法，特异性 荷尔蒙激活所必需的序列 JH酯酶的表达 3)通过凝胶延迟、DNase I足迹和甲基 与此相关的干扰特定序列或区域 JH酯酶基因 激素诱导后的结合蛋白 这些实验结果将是新的和原始的数据 JH刺激基因表达的分子机制。它的用途 在变态过程中高度表达的蛋白质，如 研究模型确保研究结果将加深我们的理解 属于并有能力颠覆一个基本的和对发展至关重要的 昆虫发育的调节点。它的意义和 结果的发生和重要性保证了结果的适用性 在广泛的无脊椎动物群体中。

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.