Molecular mechanism of gene expresion regulated by juvenile hormone : Identification and its function of Juvenile hormone receptor, responsive elements and transcription factor

我们分析了JH诱导基因CP(氰蛋白：血淋巴储存六聚体)和VG(卵黄蛋白：蝗虫卵黄蛋白)的上游调控基因结构。这两个基因的核受体超家族共有序列AGGTCA在-550bP和-1100bP之间存在反向重复。我们制备了报告基因无G盒的DNA结构，并将其限定为反向重复序列。在体外转录系统中，我们分析了这些DNA构建体以及昆虫和蝗虫脂肪体核提取液的转录活性。我们发现，JH处理动物的脂肪体核提取液通过两种昆虫的反向重复序列诱导了基因的表达。不同的基因组合和核提取液也是有效的。将这些反向重复序列与脂肪体核提取液进行凝胶印迹实验，发现JH处理后的脂肪体核提取液具有特异的移位带。这些结果提示，JH诱导的核因子可能是JH相关基因调控因子和JH受体的候选因子之一。

We have analyzed the upstream regulatory gene structure of JH inducible genes, CP (cyanoprotein : hemolymph storage hexamer) in the bean bug, Riptortus clavatus and Vg (vitellogenin : egg yolk protein in the grasshopper, Locusta migratoria). Both genes had an inverted repeat of consensus sequence AGGTCA of nuclear receptor superfamily in the region of -550bp and-1100bp. We prepared DNA constructs of a reporteer gene G-free casseteligated to the inverted repeats. We analyzed the transcriptional activities of those DNA constructs and the fat body nuclear extract of bug and locusta in the vitro transcription system. We found that the fat body nuclear extract from JH treated animal induced the gene expression using the inverted repeats from both insects. Different set of gene and nuclear extract also were effective. We have performed geldetardation assay using those inverted repeats and fat body nuclear extract, and found the specific shift band in the JH treated fat body nuclear extract. These results suggested that the nuclear factor which induced by JH are one of candidate of JH responsible gene regulatory factor and JH receptor.