REGULATION OF A DEVELOPMENTALLY CRITICAL INSECT PROTEIN

建议拨款的目的是利用荷尔蒙调节的一个 变态相关蛋白的转录机制进行研究， 保幼激素（JH）在昆虫发育过程中对基因的调控。在 粉纹夜蛾和许多其它昆虫的JH酯酶是强的， 受自身激素JH的积极调节该系统对 一项关于这种以及其他倍半萜类激素如何作用于 调节蛋白质的基因表达以及激素如何起作用， 在模式昆虫发育过程中调节基因表达， 重要的无脊椎动物。 拟议研究的具体目标和程序如下： 1)分离和测序JH酯酶cDNA和基因组克隆， 粉纹夜蛾 2)通过果蝇的体内转化或体外转化来定义 T. NI细胞系特异性序列， 所必需的激素激活的表达JH酯酶。 3)通过凝胶阻滞、DNA酶I足迹和甲基 与JH酯酶相关的干扰特异性序列或区域 在激素激活后结合蛋白质的基因。 4.启动蛋白质的纯化和鉴定， 激素激活后，与JH酯酶侧翼序列结合 基因 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。的发生和重要性 JH在广泛的无脊椎动物群体中确保了结果将 对医学无脊椎动物具有广泛的意义和适用性 重要性

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 is attractive for a study on how this, and perhaps other, sesquiterpenoid 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 Trichoplusia 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, DNAase I footprinting and methyl interference specific sequences or regions associated with the JH esterase gene which bind proteins following hormonal activation. 4.initiate purification and characterization of proteins identified as binding, after hormonal activation, to sequences flanking the JH esterase gene. 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 occurrence and importance of JH in a wide range of invertebrate groups ensures that the results will have wide significance and applicability towards invertebrates of medical importance.