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诱导基因的上游调控基因结构，分别是豆虫的cyanoprotein：血淋巴储存六聚体CP （cyanoprotein: blood淋巴储存六聚体）和蚱蜢的卵黄原蛋白Vg （vitellogenin：蛋黄蛋白）和迁徙蝗的Locusta migratoria。两个基因在-550bp和1100bp区域均存在核受体超家族共识序列AGGTCA的反向重复。我们制备了一个与反向重复序列相连接的记者基因无g cassege的DNA构建体。我们在体外转录系统中分析了这些DNA构建体和虫、蝗脂肪体核提取物的转录活性。我们发现JH处理动物的脂肪体核提取物通过两种昆虫的反向重复序列诱导基因表达。不同组的基因和核提取物也有效果。我们利用这些反向重复序列和脂肪体核提取物进行了延迟实验，发现了JH处理的脂肪体核提取物中存在特定的移位带。这些结果表明，JH诱导的核因子是JH负责基因调控因子和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.