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

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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.